Selective, lipophilic, and long-acting beta agonist monotherapeutic formulations and methods for the cosmetic treatment of adiposity and contour bulging

ABSTRACT

Provided herein are pharmaceutical and cosmetic formulations and methods for regional adiposity reduction and treatment of body contour defects such as abdominal bulging; comprising an injectable formulation, said formulation comprising: an active ingredient that consists essentially of an adipose tissue-reducing amount of one or more lipophilic long-acting selective beta-2 adrenergic receptor agonists, or salts, solvates, or polymorphs thereof; and one or more subcutaneously acceptable inactive ingredients.

CROSS-REFERENCE

This application is a continuation of U.S. patent application Ser. No.13/303,045 filed on Nov. 22, 2011 which claims the benefit of U.S.Provisional Application Ser. No. 61/417,098 filed Nov. 24, 2010, bothwhich are incorporated by reference herein in its entirety.

BACKGROUND OF THE INVENTION

Cosmetics are substances used to enhance the appearance of the humanbody. Traditionally, cosmetics have included skin-care creams, lotions,powders, perfumes, lipsticks, nail polish, eye and facial makeup, and soforth. The United States Food and Drug Administration (the FDA), whichregulates cosmetics in the United States, broadly views a cosmetic as asubstance that is intended to be applied to the human body forcleansing, beautifying, promoting attractiveness, or altering theappearance without significantly affecting the body's structure orfunctions.

Accumulation of fat stores in humans can occur unevenly in the body,which to certain individuals is considered to be a cosmetic blemish. Forexample, some persons may accumulate fat predominantly in visceral areaswhile others predominately in the subcutaneous tissue. Genderdifferences may also be apparent with women accumulating fat in thethighs and lateral buttocks and males in the waist. Women may accumulatefatty deposits of the thighs, which have a rumpled or “peau d′orange”appearance, resulting in a condition referred to as cellulite. Cellulitemay be related to skin architecture which allows subdermal fatherniation, sometimes referred to as adipose papillae.

SUMMARY OF THE INVENTION

The subject matter described herein provides cosmetic and pharmaceuticalmonotherapy formulations for the reduction of subcutaneous adiposity inhumans. Specifically, described herein are subcutaneous andtranscutaneous pharmaceutical and cosmetic monotherapy formulations andmethods of treatment for regional adiposity. Also provided herein arepharmaceutical and/or cosmetic formulations for, in certain situations,reducing regional fat deposits in a subject.

In one aspect, provided herein are injectable formulations for regionaladiposity reduction comprising: (a) active ingredient that consistsessentially of an adipose tissue-reducing amount of one or morelipophilic long-acting selective beta-2 adrenergic receptor agonists, orsalts, solvates, or polymorphs thereof; and (b) one or moresubcutaneously acceptable inactive ingredients. In another aspect,provided herein is a cosmetic method for reducing adiposity in a humanpatient comprising subcutaneously administering a formulation suitablefor subcutaneous injection comprising: (a) active ingredient thatconsists essentially of a cosmetically effective adipose tissue-reducingamount of one or more lipophilic long-acting selective beta-2 adrenergicreceptor agonists, or salts, solvates, or polymorphs thereof; and (b)one or more subcutaneously acceptable inactive ingredients. In yetanother aspect, provided herein is a cosmetic method for inducinglipolysis in adipose tissue of a human patient comprising subcutaneouslyadministering a formulation suitable for subcutaneous injectioncomprising: (a) active ingredient that consists essentially of acosmetically effective adipose tissue-reducing amount of one or morelipophilic long-acting selective beta-2 adrenergic receptor agonists, orsalts, solvates, or polymorphs thereof; and (b) one or moresubcutaneously acceptable inactive ingredients. In a further aspect,provided herein is a method for the aesthetic treatment of body contourdefects such as abdominal bulging in a human patient comprisingsubcutaneously administering a formulation suitable for subcutaneousinjection comprising: (a) active ingredient that consists essentially ofa cosmetically effective adipose tissue-reducing amount of one or morelipophilic long-acting selective beta-2 adrenergic receptor agonists, orsalts, solvates, or polymorphs thereof; and (b) one or moresubcutaneously acceptable inactive ingredients. In an aspect provided isa method for aesthetic treatment of cheek contour defect in a humanpatient by contacting a targeted fat deposit in the cheek with acomposition comprising: (a) active ingredient that consists essentiallyof a cosmetically effective adipose tissue-reducing amount of one ormore lipophilic long-acting selective beta-2 adrenergic receptoragonists, or salts, solvates, or polymorphs thereof; and (b) one or moresubcutaneously acceptable inactive ingredients. In another aspect,provided herein are compositions, formulations, methods, and systems fortreating thyroid eye disease by contacting a targeted fat deposit in theeye with a composition comprising: (a) active ingredient that consistsessentially of an adipose tissue-reducing amount of one or morelipophilic long-acting selective beta-2 adrenergic receptor agonists, orsalts, solvates, or polymorphs thereof; and (b) one or moresubcutaneously acceptable inactive ingredients.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 illustrates effects of salmeterol xinafoate on change in inguinalfat pad weight (ΔIFP) (% reduction; mean±SEM) following SC injectioninto the fat pad.

FIG. 2 shows effect of SC salmeterol xinafoate (SX) on change frombaseline back fat thickness in Yucatan minipigs determined by2D-ultrasound

DETAILED DESCRIPTION OF THE INVENTION

Adipose tissue is the primary energy storage tissue of the body. Fatcells, or adipocytes, store this energy in the form of triglycerides.Triglycerides are mobilized from fat stores to provide caloric energy tothe body through hormonal induction of triglyceride hydrolysis. Thisprocess releases free or non-esterified fatty acids and glycerol intothe blood for use by other body tissues. The breakdown of triglyceridesfrom fat store is referred to as lipolysis. Growth of new adipocytesalso occurs, which is referred to as adipogenesis. Primaryneurotransmitters that control lipolysis in the body are thecatecholamines epinephrine and norepinephrine. Adipose tissue hasbeta-1, 2, and 3 adrenergic receptors and alpha-2 adrenergic receptors.Binding of beta-adrenergic receptor agonists (“beta-adrenergicagonists”) to beta-adrenergic (“beta”) receptors in adipose tissueresults in adipocyte lipolysis. Beta-adrenergic receptor activation alsoinhibits adipogenesis. In humans, beta-2 receptors are the most abundanton fat cell surfaces and the primary mediator of beta-adrenergicreceptor-stimulated lipolysis. Stimulation of lipolysis bybeta-adrenergic agonists is mediated by adenylate cyclase and increasedformation of cyclic adenosine monophosphate (cyclic AMP, cAMP).

Long-acting beta-2 adrenergic receptor agonists, such as salmeterol andformoterol, reduce regional fat deposits or adipose tissue regions bybinding to beta receptors, resulting in adipocyte lipolysis. The use oflong-acting beta-2 adrenergic receptor agonists, however, carries withit possible side effects. For example, use of long-acting beta-2adrenergic receptor agonists may result in cardiovascular problems suchas angina, hypertension or hypotension, tachycardia, palpitations, andarrhythmias. Thus, while long-acting beta-2 adrenergic receptor agonistsmay reduce regional fat deposits and adipose tissue regions they mayalso cause increased heart rate and palpitations.

It has been found that certain lipophilic long-acting selective beta-2adrenergic agonists administered subcutaneously in appropriate amountsreduce regional fat deposits with limited systemic exposure compared toother long-acting beta-2 adrenergic agonists. One possible reason forthis result is that the lipophilic nature of certain long-acting beta-2adrenergic receptor agonists allows selective partitioning into theadipose tissue relative to plasma. The lipophilicity of certainlong-acting beta-2 adrenergic receptor agonist into the subcutaneousadipose tissue contributes, at least in part, to providing relativelylow levels of the agonist systemically. Appropriate amounts of thelipophilic long-acting beta-2 adrenergic receptor agonists describedherein administered via subcutaneous or transcutaneous injection mayprovide therapeutic effectiveness in reducing regional fat depositsand/or adipose tissue with a reduced risk of producing cardiovascularside effects. Indeed, the formulations described herein are prepared tobe administered subcutaneously or transcutaneously, and notsystemically. Accordingly, the formulations and methods of treatmentdescribed herein are designed to reduce subcutaneous adipose tissuewhich is distinct from visceral (systemic) fat.

GLOSSARY OF CERTAIN TERMINOLOGY

“Cosmetically effective” as used herein, refers to a sufficient amountof an agent (e.g., a selective, long-acting, and long-acting beta-2agonist) which will improve the cosmetic appearance at the localizedsite of treatment. A “cosmetically effective” amount of a selective,lipophilic, and long-acting beta-2 agonist is an amount effective toachieve a cosmetically desirable improvement. It is to be understoodthat a “cosmetically effective” amount can vary from subject to subject,due to numerous factors including for example age, weight, generalcondition of the subject, the condition being treated, the severity ofthe condition being treated, and the judgment of the prescribingphysician.

As used herein, an “aesthetic treatment” or “cosmetic treatment” refersto any treatment carried out for the purposes of improving the visualappearance. A method for aesthetic treatment refers to a method ofimproving the visual appearance at the localized site of treatment byproviding an effective amount of an agent (e.g., a selective,long-acting, and long-acting beta-2 agonist) which will improve thecosmetic appearance at the localized site of treatment. As used herein,the aesthetic treatment of a body contour defect refers to any treatmentcarried out to achieve a more natural or cosmetically desirable bodyshape.

As used herein an “adipose tissue-reducing” amount refers to asufficient amount of the lipophilic long-acting beta-2 adrenergicreceptor agonist needed to reduce adipose tissue. It is to be understoodthat the amount sufficient to decrease the adipose tissue will vary fromsubject to subject due to variation in metabolism of the lipophiliclong-acting beta-2 adrenergic receptor agonist, with age, weight,general condition of the subject, the severity of the condition beingtreated, and the judgment of the prescribing physician.

As used herein, “Thyroid Eye Disease” or “Graves' Opthamolopathy” or“Thyroid-associated orbitopathy” or “Grave's orbitopathy” refers to aneye condition that is characterized by one or more of: swelling in theorbital tissues; protrusion of one or both eyeballs—also known asexophthalmos; proptosis, in which the eyes appear to bulge outward.Without being limited by theory, it is believed the clinical symptomsand signs of Thyroid Eye Disease can be explained mechanically by theincrease in tissue volume evident within the bony orbit. The expandedorbital tissues cause forward displacement of the globe and impairmentof venous and lymphatic outflow from the orbit. These changes, combinedwith the local production of cytokines and other mediators ofinflammation, result in proptosis, periorbital edema, conjunctivalerythema and/or chemosis.

A formulation that is “substantially free” of a specific compound orsubstance contains an amount that is equal to or less than a trace ortherapeutically or cosmetically insignificant amount of the specificcompound or substance. For example, a formulation that is “substantiallyfree” of a glucocorticosteroid contains an amount that is equal to orless than a trace of therapeutically or cosmetically insignificantamount of glucocorticosteroids.

As described herein a “reduced or minimized risk of producingcardiovascular side effects” amount refers to an amount of thelipophilic long-acting beta-2 adrenergic receptor agonist used whichdoes not result in clinically significant cardiovascular side effects.It is to be understood that the amount used will vary from subject tosubject due to variation in metabolism of the lipophilic long-actingbeta-2 adrenergic receptor agonist, with age, weight, general conditionof the subject, the severity of the condition being treated, and thejudgment of the prescribing physician.

“Plasma concentration” refers to the concentration of a substance suchas a therapeutic agent, in blood plasma of a subject. It is understoodthat the plasma concentration of a therapeutic agent may vary many-foldbetween subjects, due to variability with respect to metabolism oftherapeutic agents. In accordance with one aspect, the plasmaconcentration of a long-acting beta-2 adrenergic receptor agonist variesfrom subject to subject. Likewise, in some embodiments, values such asmaximum plasma concentration (C_(max)) or time to reach maximum plasmaconcentration (T_(max)), or total area under the plasma concentrationtime curve (AUC) varies from subject to subject. Due to thisvariability, the amount necessary to constitute “a therapeuticallyeffective amount” of a lipophilic long-acting beta-2 adrenergic receptoragonist varies from subject to subject. It is understood that in someembodiments, when mean plasma concentrations are disclosed for apopulation of subjects, these mean values include substantial variation.

“Pharmacodynamics” refers to the factors that determine the biologicresponse observed relative to the concentration of drug at a site ofaction.

“Pharmacokinetics” refers to the factors that determine the attainmentand maintenance of the appropriate concentration of drug at a site ofaction.

A “measurable plasma concentration” or “measurable plasma concentration”describes the blood plasma or blood plasma concentration, typicallymeasured in mg, μg, or ng of therapeutic agent per mL, dL, or L of bloodplasma, of a therapeutic agent that is absorbed into the bloodstreamafter administration. One in the field would be able to measure theplasma concentration or plasma concentration of a lipophilic long-actingbeta-2 adrenergic receptor agonist.

Some embodiments comprise optically pure isomers of the lipophilicbeta-adrenergic agonist(s), which improve lipolysis and adipogenesisinhibition and reduce the risk of producing potential cardiovascularside effects. In some embodiments, these optically pure isomers allowformulations comprising larger loadings of an active ingredient, forexample, by eliminating one or more isomers with no physiologicaleffect, a lesser a physiological effect, a negative effect, and/or anundermined physiological effect. Removing the undesired bounds of aracemic mixture isolates the active isomer, or eutomer, thereby allowingmore eutomer to be loaded in a give formulation by removing the inactivecomponents.

Two stereogenic centers in a molecule generally generate twodiastereomers, referred to herein as (R*,R*) and (R*,S*), and theirenantiomers. Diastereomers are stereoisomers that are not enantiomers,that is, the mirror image of one diastereomer is not superimposable onanother diastereomer. Enantiomers are stereoisomers that are mirrorimages of each other. A racemate is a 1:1 mixture of enantiomers. Theenantiomers of the (R*,R*) diastereomers are referred to as the (R,R)and (S,S) enantiomers, which are mirror images of each other andtherefore share some chemical and physical properties, for examplemelting points. Similarly, the (R,S) and (S,R) isomers are enantiomersof the (R*,S*) enantiomer. For example, some embodiments compriseoptically pure isomers of lipophilic beta-2 agonists, for example,(R)-salmeterol.

Additionally, in some embodiments, long-acting selective beta-2 agonistsare lipophilic, thereby providing a pharmaceutical and/or cosmeticformulation with activity in fat tissue. In some embodiments, thelipophilic agonist is salmeterol. In further embodiments, thelipophilicity of salmeterol provides prolonged exposure to the adiposetissue. In some embodiments, the agent is not salmeterol, but has asimilar lipophilicity to salmeterol.

Salmeterol has high lipid solubility, compared to other long-actingbeta-2 adrenergic receptor agonists, such as for example, formoterol,which extends its residence time in the adipose tissue and/or in one ormore adipose cells. Some embodiments of the subcutaneous formulationcomprise a highly lipophilic beta-adrenergic agonist, which reduces oreliminates the need for a sustained or controlled release carrier due topartitioning and sequestration in the adipose tissue thereby prolongingthe treatment effect. In some embodiments, lipophilic beta-adrenergicagonists with an oil-water partition coefficient of at least about 1000or at least about 10,000 to 1 are used. For example, salmeterol is atleast 10,000 times more lipophilic than albuterol, a short-actinghydrophilic beta-adrenergic agonist.

A “treatment period” is defined as the period of time the patient isunder a physician's care or direction, which may vary from patient topatient, and may be dependent on metabolism of the lipophiliclong-acting beta-2 adrenergic receptor agonist administered to thepatient, age, weight, general condition of the subject, the severity ofthe condition being treated, and the judgment of the prescribingphysician. In some embodiments, the treatment period comprises between 1week and 52 weeks, longer than 52 weeks, or any amounts of weeks between1 and 52.

A “weekly dose” is the total amount of active ingredient administered toa patient during a single week. For example, in situations with morethan a single administration occurs during a week, the weekly dose isthe total amount of active ingredient provided to the patient in eachadministration that occurs during the week.

A “periodic dose” is the frequency at which a dose is administered to apatient during a period.

A “single session dose” is the total amount of active ingredientadministered to a patient during a single visit for treatment by ahealthcare professional or, in situations of self-administration, asingle session dose is the total amount of active ingredientadministered to the patient by self-administration in a single session.

In some embodiments, a single session dose is divided into smalleramounts and administered to a patient in one or more “sub-doses.” Insome embodiments, each “sub-dose” is subcutaneously delivered to apatient by injection, e.g., using a syringe or is administered to thepatient transcutaneously.

The phrases “patient” and “subject” are used interchangeably herein. Insome embodiments, the patient or subject is a human. In further oradditional embodiments, the patient or subject is an animal. In someembodiments, the animal is a human, a common household pet, includingfor example a cat or a dog, or a species of the animal kingdom. In someembodiments, the patient is a non-murine animal.

Lipophilic, Selective, and Long-Acting Beta-2 Adrenergic ReceptorAgonists

In one aspect, provided herein are pharmaceutical and/or cosmeticformulations suitable for subcutaneous or transcutaneous administrationand methods of treatment comprising subcutaneously or transcutaneouslyadministering to a patient a pharmaceutical and/or cosmetic formulation(including all of the methods of treatment described herein). In someembodiments, the pharmaceutical and/or cosmetic formulation comprises aninjectable formulation for regional adiposity reduction consistingessentially of: (a) active ingredient that consists essentially of anadipose tissue-reducing amount of one or more lipophilic long-actingselective beta-2 adrenergic receptor agonists, or salts, solvates, orpolymorphs thereof; and (b) one or more subcutaneously acceptableinactive ingredients. In further or additional embodiments, theformulations provided herein comprise an injectable formulation forregional adiposity reduction consisting of: (a) active ingredient thatconsists essentially of an adipose tissue-reducing amount of one or morelipophilic long-acting selective beta-2 adrenergic receptor agonists, orsalts, solvates, or polymorphs thereof; and (b) one or moresubcutaneously acceptable inactive ingredients. In further or additionalembodiments, the formulations provided herein comprise an injectableformulation for regional adiposity reduction comprising: (a) activeingredient that consists essentially of an adipose tissue-reducingamount of one or more lipophilic long-acting selective beta-2 adrenergicreceptor agonists, or salts, solvates, or polymorphs thereof; and (b)one or more subcutaneously acceptable inactive ingredients. In specificembodiments of the methods and formulations described herein, theinjectable formulations are substantially free of glucocorticosteroids.

In some embodiments, provided herein are pharmaceutical and/or cosmeticformulations, and methods of treatment, comprising administration of thepharmaceutical and/or cosmetic formulations to a patient, wherein theformulation is suitable for subcutaneous administration. In further oradditional embodiments, the pharmaceutical and/or cosmetic formulationis suitable for transcutaneous administration.

In some embodiments, the pharmaceutical and/or cosmetic formulationcomprises a lipophilic long-acting selective beta-2 adrenergic receptoragonist, or a salt, optical isomer, racemate, solvate, or polymorphthereof. For example, in some embodiments, the lipophilic long-actingselective beta-2 adrenergic receptor agonist is salmeterol (includingvarious salt forms such as xinafoate). In further or additionalembodiments the lipophilic long-acting selective beta-2 adrenergicreceptor agonist is formoterol (including various salt forms such asfumarate and furoate).

In other embodiments, the pharmaceutical and/or cosmetic formulationconsists essentially of a lipophilic long-acting selective beta-2adrenergic receptor agonist, or a salt, optical isomer, racemate,solvate, or polymorph thereof

In one aspect, provided herein are pharmaceutical and/or cosmeticformulations suitable for subcutaneous or transcutaneous administrationcomprising a lipophilic long-acting selective beta-2 adrenergic receptoragonist, including, for example, salmeterol or2-(hydroxymethyl)-4-{1-hydroxy-2-[6-(4-phenylbutoxy)hexylamino]ethyl}phenol, or its salts, optical isomers, racemates,solvates or polymorphs thereof and when used in the appropriate amountsand administered transcutaneously or subcutaneously, provides atherapeutic effect for reducing regional fat deposits and/or adiposetissue with limited systemic exposure, and consequently, a reduced riskof producing cardiovascular side effects. In one embodiment is asubcutaneous or transcutaneous preparation for the reduction of adiposetissue and/or the reduction in regional fat deposits comprising anadipose tissue-reducing amount of a lipophilic long-acting selectivebeta-2 adrenergic agonist wherein the formulation does not result inhigh systemic levels when administered subcutaneously ortranscutaneously. In another embodiment, the lipophilic long-actingselective beta-2 adrenergic receptor agonist is salmeterol. In yetanother embodiment, the agonist is a polymorph of salmeterol, such asfor example, polymorph I and II. Such subcutaneous or transcutaneouspreparations provide the required tissue concentration of salmeterolneeded to reduce adipose tissue and/or reduce regional fat deposits witha minimized or reduced risk of producing the side effects typicallyassociated with the administration of beta-2 adrenergic receptoragonists, including other long-acting beta-2 adrenergic receptoragonists. Additionally, the use of salmeterol in a subcutaneous ortranscutaneous preparation provides therapeutically effective dosageswithout producing relatively high systemic levels found when using otherlong-acting beta-2 adrenergic receptor agonists, such as for example,formoterol.

In another aspect, provided herein are pharmaceutical and/or cosmeticformulations suitable for subcutaneous or transcutaneous administrationand methods of treatment comprising subcutaneously or transcutaneouslyadministering to a patient a pharmaceutical and/or cosmetic formulation(including all of the methods of treatment described herein) wherein thepharmaceutical and/or cosmetic formulation comprises a beta-2 adrenergicreceptor agonist.

In some embodiments, provided herein are pharmaceutical and/or cosmeticformulations and methods of treatment comprising a beta-2 adrenergicreceptor agonist, or a salt, optical isomer, racemate, solvate, orpolymorph thereof, that is characterized by at least one of thefollowing properties: lipophilic; selective for the beta-2 adrenergicreceptor; and long-acting. In some embodiments, the beta-2 adrenergicreceptor agonist is selective for the beta-2 adrenergic receptor. Infurther or additional embodiments, the beta-2 adrenergic receptoragonist is lipophilic. In further or additional embodiments, the beta-2adrenergic receptor agonist is long-acting.

In some embodiments, the beta-2 adrenergic receptor agonist isbambuterol, bitolterol, broxaterol, carbuterol, carmoterol, clenbuterol,ibuterol, sulfonterol, isoproterenol, trimetoquinol, formoterol,desformoterol, hexoprenaline, ibuterol, indacaterol, isoetharine,isoprenaline, isoproterenol, levalbuterol, metaproterenol, picumeterol,pirbuterol, procaterol, reproterol, rimiterol, salbutamol, salmeterol,sulfonterol, terbutaline, trimetoquinol, tulobuterol, TA-2005(8-hydroxy-5-((1R)-1-hydroxy-2-(N-((1R)-2-(4-methoxyphenyl)-1-methylethyl-)amino)ethyl)-carbostyrilhydrochloride), QAB-149 (Novartis), TA-2005, GSK-159797, or GSK-642444,or a salt, optical isomer, racemate, solvate, or polymorph thereof

In some embodiments, the beta-2 adrenergic receptor agonist islong-acting and is selected from salmeterol, formoterol, bambuterol, orclenbuterol. In further or additional embodiments, the beta-2 adrenergicreceptor agonist is a specific type of long-acting that is ultralong-acting. In some embodiments, the ultra long-acting beta-2adrenergic receptor agonist is selected from indacaterol, carmoterol,QAB-149, CHF-4226, TA-2005, GSK-159797, and GSK-642444.

In some embodiments, provided herein are pharmaceutical and/or cosmeticformulations comprising an active ingredient consisting essentially ofan adipose tissue-reducing amount of a lipophilic long-acting selectivebeta-2 adrenergic receptor agonist or a salt, optical isomer, racemate,solvate, or polymorph thereof and at least one subcutaneously ortranscutaneously acceptable inactive ingredient. In one embodiment, thelipophilic long-acting selective beta-2 adrenergic receptor agonistselectively partitions into adipose tissue relative to plasma.

In another embodiment, the lipophilic long-acting selective beta-2adrenergic receptor agonist is salmeterol(±2-(hydroxymethyl)-4-[1-hydroxy-2-[6-(4-phenylbutoxy)hexylamino]ethyl]-phenol,CAS Reg. No. 94749-08-3, shown below as compound 1).

In other embodiments, the lipophilic, long-acting, selective beta-2agonist is a polymorph of salmeterol. In a further embodiment, thepolymorph is polymorph I or II. In further embodiments, the formulationuses a mixture of salmeterol polymorphs. In yet another embodiment, thesalt of the lipophilic long-acting selective beta-2 adrenergic receptoragonist is a xinafoate salt. In some embodiments, the lipophiliclong-acting selective beta-2 adrenergic receptor agonist is salmeterolxinafoate.

Some embodiments provide adrenergic modulation through the use ofpharmaceutical and/or cosmetic compositions comprising an adiposetissue-reducing and lipophilic, long-acting selective beta-2 receptoragonist active ingredient administered subcutaneously ortranscutaneously, either alone (as single agent therapy), or incombination with at least a second active ingredient (as a combinationtherapy). Thus, in some embodiments, the pharmaceutical and/or cosmeticformulations (and corresponding methods of treatment provided herein)consist essentially of a lipophilic, long-acting, selective beta-2agonist, for example, salmeterol, physiologically acceptable salts,optical isomers, racemates, solvates, polymorphs, or combinationsthereof, wherein the formulation is suitable for subcutaneous ortranscutaneous administration. In further or additional embodiments, thepharmaceutical and/or cosmetic formulation comprises a lipophilic,long-acting, selective beta-2 agonist, for example, salmeterol,physiologically acceptable salts, optical isomers, racemates, solvates,polymorphs, or combinations thereof, and at least a second activeingredient, wherein the formulation is suitable for subcutaneous ortranscutaneous administration.

Lipophilic, long-acting, selective beta-2 agonists, for example,salmeterol are used in some embodiments. In other embodiments, salts,optical isomers, racemates, polymorphs, and/or solvates of beta-2agonists have the desired activity and are accordingly provided forherein. Unless otherwise specified, references to an active ingredient,for example, to salmeterol, include the compound itself as well as aphysiologically acceptable analogs, salts, optical isomers, racemates,polymorphs, solvates, or combinations thereof.

In some embodiments, salmeterol is used in the compositions and methodsdescribed herein. Depending on the tissue, salmeterol may exhibitpartial agonist activity, which is believed to reduce receptordesensitization and may limit arrestin signaling leading to lessreceptor down-regulation. In some embodiments, salmeterol is present asa physiologically acceptable salt, optical isomer, racemate, solvate,and/or polymorph thereof. Suitable physiologically acceptable salts ofsalmeterol include, but are not limited to acid addition salts derivedfrom inorganic and organic acids, such as the hydrochloride,hydrobromide, sulfate, phosphate, maleate, tartrate, citrate, benzoate,4-methoxybenzoate, 2-hydroxybenzoate, 4-hydroxybenzoate,4-chlorobenzoate, p-toluenesulphonate, methanesulphonate, ascorbate,salicylate, acetate, fumarate, succinate, lactate, glutarate, gluconate,tricarballylate, hydroxynaphthalenecarboxylate,1-hydroxy-2-naphthalenecarboxylate, 3-hydroxy-2-naphthalenecarboxylate,oleate, combinations thereof, and the like. In some embodiments,salmeterol is provided as the 1-hydroxy-2-naphthalene carboxylate salt(hydroxynaphthoate, also known as xinafoate).

Dosing

In some embodiments, long-term exposure of adipose tissue to adrenergicagents, particularly beta-adrenergic receptor agonists, results inreceptor desensitization through receptor phosphorylation andsequestration. These effects limit the ability of an adrenergicmodulating composition to treat adipose tissue and result intachyphylaxis, a condition in which the body experiences a rapidlydecreasing response to the agonist following administration of theinitial doses, to the desired lipolytic and anti-adipogenesis effect.Consequently, in certain situations with long-term exposure of adiposetissue to beta-adrenergic receptor agonists, the therapeutic effect withthe beta-adrenergic receptor agonists is short-lived.

Repeated administration of short-acting beta-2 agonists often result intachyphylaxis. However, salmeterol, exhibits partial beta-2 receptoragonist activity in some systems that may reduce the desensitizationthat occurs with continuous exposure of adipocytes to full adrenergicreceptor agonists. Compared with short-acting beta-2 agonists, lipolysisalso occurs for a longer time after administration because lipophilic,long-acting selective beta-2 agonists have longer half-lives. Thecombination of longer half-lives and activities reduces the requiredfrequency and total dosage of administration of the pharmaceuticaland/or cosmetic compositions provided herein. Consequently, in someembodiments, daily administration or more than once daily administrationof the composition is not required. In some embodiments, provided hereinare subcutaneously or transcutaneously administered adiposetissue-reducing lipophilic, long-acting selective beta-2 agonists whichexhibit greater selectivity for beta-2 receptors, permittingsubstantially similar therapeutic effects with less selective beta-2agonists at a lower dosage and/or less frequent dosage. Further the moreselective beta-2 activity can limit cardiac and other systemic sideeffects, which in the case of cardiac side effects, is often induced bybeta-1 receptor stimulation in the heart. In some embodiments, providedare subcutaneous or transcutaneous formulations of lipophilic,long-acting beta-2 agonists which provide selectivity for beta-2receptors while reducing the risk of producing cardiac or systemic sideeffects.

Provided herein are pharmaceutical and/or cosmetic formulations that aresuitable for subcutaneous or transcutaneous administration.

In some embodiments, the pharmaceutical and/or cosmetic formulationsprovided herein are suitable for subcutaneous injection, and provide fora volume of up to about 20 mL (including, e.g., about 0.1 mL, about 0.3mL, about 0.5 mL, about 0.7 mL, about 1.0 mL, about 1.1 mL, about 1.5mL, about 2 mL, about 2.5 mL, about 3 mL, about 3.5 mL, about 4 mL,about 5 mL, about 6 mL, about 7 mL, about 8 mL, about 9 mL, or any othervolume from about 0.1 mL to about 20 mL) of an excipient compatible withsubcutaneous administration. In some embodiments, the volume forinjection is provided in a range that is between about 0.1 mL to about20 mL, about 0.2 mL to about 15 mL, about 0.3 mL to about 10 mL, about0.4 mL to about 7 mL, about 0.5 mL to about 5 mL, about 0.6 mL to about4 mL, about 0.7 mL to about 3 mL, about 0.8 mL to about 2 mL, or about 1mL. In some embodiments, the excipient concentration is kept below 1%(e.g., about 0.05%, about 0.2%, about 0.3%, about 0.4%, about 0.5%,about 0.6%, about 0.8%, or any other concentration from about 0.05% toless than about 1%. In specific embodiments, the excipient concentrationis between about 0.01% to about 1%, about 0.05% to about 0.9%, about0.2% to about 0.8%, about 0.3% to about 0.7%, about 0.4% to about 0.6%,or about 0.5%.

Periodic Dosing Schedule

Another aspect of the formulations and methods of treatment providedherein is a periodic dosing schedule. A periodic dose is the frequencyat which a single session dose is administered to a patient during aperiod. For example, in some embodiments, the periodic dose is once perweek, and hence in these situations a patient will receive a singlesession dose once per week. In further or additional embodiments, theperiodic dose is 2-7 times per week (including any interval between 2and 7), 3-6 times per week (including any interval between 3 and 6), or4-5 days per week. In some embodiments, the periodic dose is 1-4 timesper month (including any interval between 1 and 4), 2-3 times per month,or once or twice per month. In some embodiments, the periodic dose is1-52 times per year (including any interval between 1 and 52).

In some situations where the periodic dose is twice per week, thepatient will receive the single session doses in two separate halves(that are about equal or unequal) during the week. Similarly, in somesituations where the periodic dose is seven times per week, the amountof active ingredient administered to the patient for each single sessiondose compared to what is provided herein is divided by seven. As anotherexample, in certain situations where the periodic dose is once permonth, the patient will receive a single session dose per month at fourtimes the amount that is provided herein.

Session Dose

An additional aspect of the formulations and methods of treatmentprovided herein is session dosing. A single session dose is the totalamount of active ingredient administered to a patient during a singlevisit for treatment by a healthcare professional or, in situations ofself-administration, it is the amount of active ingredient administeredto the patient by self-administration in a session. The single sessiondose amounts provided herein are based on a once per week periodic dose,and the amounts can be adjusted for a different periodic dose than onceper week as provided herein.

In one aspect, including certain methods of treatment comprisingadministration of the pharmaceutical and/or cosmetic formulationsdescribed herein, provided are pharmaceutical and/or cosmeticformulations wherein a lipophilic long-acting selective beta-2adrenergic receptor agonist is provided in a single session dose that isless than about 20 μg of the lipophilic long-acting selective beta-2adrenergic receptor agonist, or a salt, optical isomer, racemate,solvate, or polymorph thereof. In some embodiments, the lipophilic,long-acting selective beta-2 agonist is salmeterol, or a salt, opticalisomer, racemate, solvate, or polymorph thereof. In further embodiments,the lipophilic, long-acting selective beta-2 agonist is salmeterolxinafoate. In further or additional embodiments, the salmeterolxinafoate is administered in a periodic dose that is once per week, andis administered to a patient in single session dose as provided forherein.

In some embodiments, the single session dose of salmeterol xinafoate isadministered once per week in an amount between about 20 μg and about 5ng, between about 20 μg and about 25 ng, between about 20 μg and about50 ng, between about 20 μg and about 75 ng, between about 20 μg andabout 100 ng, between about 20 μg and about 125 ng, between about 20 μgand about 150 ng, between about between about 20 μg and about 175 ng,between about 20 μg and about 200 ng, between 20 μg and about 225 ng,between about 20 μg and about 250 ng, between about 20 μg and about 275ng, between about 20 μg and about 300 ng, between about 20 μg and about325 ng, between about 19 μg and about 350 ng, between about 19 μg andabout 375 ng, between about 18 μg and about 400 ng, between about 18 μgand about 425 ng, between about 18 μg and about 450 ng, between about 18μg and about 475 ng, between about 18 μg and about 500 ng, between about17 μg and about 525 ng, between about 17 μg and about 550 ng, betweenabout 17 μg and about 575 ng, between about 17 μg and about 600 ng,between about 17 μg and about 625 ng, between about 17 μg and about 650ng, between about 16 μg and about 675 ng, between about 15 μg and about700 ng, between about 14 μg and about 725 ng, between about 13 μg andabout 750 ng, between about 13 μg and about 775 ng, between about 13 μgand about 800 ng, between about 12 μg and about 825 ng, between about 11μg and about 850 ng, between about 10 μg and about 875 ng, between about9 μg and about 900 ng, between about 8 μg and about 920 ng, betweenabout 7 μg and about 940 ng, between about 6 μg and about 950 ng,between about 5 μg and about 960 ng, between about 4 μg and about 980ng, or between about 3 μg and about 1 μg. In some embodiments, alipophilic, long-acting selective beta-2 agonist to be administered issalmeterol and an adipose tissue-reducing amount of salmeterol to beadministered is less than about 5 μg per week.

In specific embodiments, provided are pharmaceutical and/or cosmeticformulations that are formulated to provide a daily dose of alipophilic, long-acting selective beta-2 agonist. In some embodiments, alipophilic, long-acting selective beta-2 agonist to be administered issalmeterol and an adipose tissue-reducing amount of salmeterol to beadministered is about 0.001 μg/day to about 1000 μg/day, e.g., about 0.1μg/day to about 100 μg/day, about 1 μg/day to about 100 μg/day, about 10μg/day to about 100 μg/day, about 50 μg/day to about 100 μg/day, or anyother dose of salmeterol from about 0.001 μg/day to about 1000 μg/day.In some embodiments, a lipophilic, long-acting selective beta-2 agonistto be administered is salmeterol and an adipose tissue-reducing amountof salmeterol to be administered is a daily dose that provides less thanabout 5 μg per week.

Sub-Dosing

In certain situations, the single session dose is administered to thepatient in sub-doses (e.g., by subcutaneous injection, transcutaneousapplication, or otherwise). Accordingly, in another aspect, includingcertain methods of treatment comprising administration of thepharmaceutical and/or cosmetic formulations described herein, providedare pharmaceutical and/or cosmetic formulations wherein a lipophiliclong-acting selective beta-2 adrenergic receptor agonist is provided inat least two sub-doses. In some embodiments, all of the sub-doses areprovided to a patient in a single session during a single week. Infurther or additional embodiments, including certain methods oftreatment comprising administration of the pharmaceutical and/orcosmetic formulations described herein, the active ingredient isprovided in at least two sub-doses whereby all of the sub-doses areprovided to a patient in a single session during a single week. In stillfurther embodiments, at least at least two sub-doses are provided to apatient.

In some embodiments, one or more sub-dose is provided to a patientwherein each sub-dose is a single injection of a fluid comprising alipophilic long-acting selective beta-2 adrenergic receptor agonist. Forexample, in some embodiments, provided herein are pharmaceutical and/orcosmetic formulations and methods of treatment comprising administrationof a pharmaceutical and/or cosmetic formulation wherein a lipophiliclong-acting selective beta-2 adrenergic receptor agonist is provided toa patient in about a single sub-dose, at least about two sub-doses, atleast about three sub-doses, at least about four sub-doses, at leastabout five sub-doses, at least about six sub-doses, at least about sevensub-doses, at least about eight sub-doses, at least about ninesub-doses, at least about 10 sub-doses, at least about 11 sub-doses, atleast about 12 sub-doses, at least about 13 sub-doses, at least about 14sub-doses, at least about 15 sub-doses, at least about 16 sub-doses, atleast about 17 sub-doses, at least about 18 sub-doses, at least about 19sub-doses, at least about 20 sub-doses, at least about 21 sub-doses, atleast about 22 sub-doses, at least about 23 sub-doses, at least about 24sub-doses, at least about 25 sub-doses, at least about 26 sub-doses, atleast about 27 sub-doses, at least about 28 sub-doses, at least about 29sub-doses, at least about 30 sub-doses, at least about 31 sub-doses, atleast about 32 sub-doses, at least about 33 sub-doses, at least about 34sub-doses, at least about 35 sub-doses, or more than about 35 sub-doses.In further or additional embodiments, a lipophilic long-acting selectivebeta-2 adrenergic receptor agonist is provided to a patient in about a 1to about 35 sub-doses, about 2 to about 32 sub-doses, in about 3 toabout 30 sub-doses, in about 5 to about 28 sub-doses, in about 7 toabout 27 sub-doses, in about 9 to about 26 sub-doses, in about 11 toabout 25 sub-doses, in about 12 to about 24 sub-doses, in about 14 toabout 23 sub-doses, in about 16 to about 22 sub-doses, in about 17 toabout 21 sub-doses, or in about 18 to about 20 sub-doses.

In some embodiments, each sub-dose is administered to a patient in anequal amount. For example, in some situations where the single sessiondose is about 20 μg of salmeterol xinafoate that is delivered to thepatient in 20 sub-doses, each sub-dose contains about 1 μg of salmeterolxinafoate. In other situations, the single session dose is about 500 ngand is delivered to the patient in 22 sub-doses and each sub-dosecontains about 22 or 23 ng of salmeterol xinafoate. Still in furthersituations, a prescribing physician may administer, or the patient mayself-administer, sub-doses in amounts that are not equal but vary inamount with respect to each sub-dose that is administered.

In some embodiments, at least two sub-doses of salmeterol xinafoate, asdescribed herein, are administered to a patient in a single session dosevia subcutaneous injection to the abdominal region of the patient. Insome of these embodiments, each sub-dose is applied to a patient about2-6 cm away from a closest second sub-dose. In further embodiments, eachsub-dose is applied to a patient about 4 cm away from a closest secondsub-dose.

In some embodiments, a sub-dose is administered, for example bysubcutaneous or transcutaneous injection, to areas of non-visceral fatdeposits on a subject, including for example subcutaneous fat. In someembodiments for which the formulations described herein are usefulinclude, but are not limited to, the inside region of the knees, themiddle to upper area of the upper arm, including the tricep area, thesubmental area, including the area under the chin, for example thewattle (which is understood to refer to the fleshy fold of skin in thesubmental area of a patient), the abdomen, the hips, the inner thigh,the outer thigh, the buttocks, an upper arms region of the patient, thelower back, upper back and the chest.

In certain embodiments, provided are pharmaceutical and/or cosmeticformulations and methods of treatment comprising the pharmaceuticaland/or cosmetic formulations that are formulated to provide a sub-doseof salmeterol xinafoate between about 20 μg and about 1 ng, betweenabout 20 μg and about 2 ng, between about 20 μg and about 3 ng, betweenabout 20 μg and about 4 ng, between about 20 μg and about 5 ng, betweenabout 20 μg and about 6 ng, between about 20 μg and about 7 ng, betweenabout 15 μg and about 8 ng, between about 10 μg and about 9 ng, betweenabout 5 μg and about 10 ng, between about 1 μg and about 12 ng, betweenabout 900 ng and about 14 ng, between about 800 ng and about 16 ng,between about 700 ng and about 18 ng, between about 600 ng and about 20ng, between about 550 ng and about 22 ng, between about 500 ng and about24 ng, between about 450 ng and about 26 ng, between about 400 ng andabout 28 ng, between about 350 ng and about 30 ng, between about 300 ngand about 32 ng, between about 250 ng and about 34 ng, between about 200ng and about 36 ng, between about 150 ng and about 38 ng, between about125 ng and about 40 ng, between about 100 ng and about 42 ng, betweenabout 90 ng and about 44 ng, between about 80 ng and about 46 ng,between about 75 ng and about 48 ng, between about 70 ng and about 50ng, between about 69 ng and about 51 ng, between about 68 ng and about52 ng, between about 67 ng and about 53 ng, between about 66 ng andabout 54 ng, between about 65 ng and about 55 ng, between about 64 ngand about 56 ng, between about 63 ng and about 57 ng, between about 62ng and about 58 ng, between about 61 ng and about 59 ng, or about 60 ng.

In some embodiments, provided herein are pharmaceutical and/or cosmeticformulations, and methods of treatment comprising administration of thepharmaceutical and/or cosmetic formulations, that are formulated toprovide a sub-dose of salmeterol xinafoate that is equal to or less thanabout 20 μg, equal to or less than about 19 μg, equal to or less thanabout 18 μg, equal to or less than about 17 μg, equal to or less thanabout 16 μg, equal to or less than about 15 μg, equal to or less thanabout 14 μg, equal to or less than about 13 μg, equal to or less thanabout 12 μg, equal to or less than about 11 μg, equal to or less thanabout 10 μg, equal to or less than about 9 μg, equal to or less thanabout 8 μg, equal to or less than about 7 μg, equal to or less thanabout 6 μg, equal to or less than about 5 μg, equal to or less thanabout 4 μg, equal to or less than about 3 μg, equal to or less thanabout 2 μg, equal to or less than about 1 μg, equal to or less thanabout 975 ng, equal to or less than about 950 ng, equal to or less thanabout 925 ng, equal to or less than about 900 ng, equal to or less thanabout 875 ng, equal to or less than about 850 ng, equal to or less thanabout 825 ng, equal to or less than about 800 ng, equal to or less thanabout 775 ng, equal to or less than about 750 ng, equal to or less thanabout 725 ng, equal to or less than about 700 ng, equal to or less thanabout 675 ng, equal to or less than about 650 ng, equal to or less thanabout 625 ng, equal to or less than about 600 ng, equal to or less thanabout 575 ng, equal to or less than about 550 ng, equal to or less thanabout 525 ng, equal to or less than about 500 ng, equal to or less thanabout 475 ng, equal to or less than about 450 ng, equal to or less thanabout 425 ng, equal to or less than about 400 ng, equal to or less thanabout 375 ng, equal to or less than about 350 ng, equal to or less thanabout 325 ng, equal to or less than about 300 ng, equal to or less thanabout 275 ng equal to or less than about 250 ng, equal to or less thanabout 225 ng, equal to or less than about 200 ng, equal to or less thanabout 175 ng, equal to or less than about 150 ng, equal to or less thanabout 125 ng, equal to or less than about 100 ng, equal to or less thanabout 75 ng, or equal to or less than about 50 ng, or equal to or lessthan about 25 ng, equal to or less than about 20 ng, equal to or lessthan about 15 ng, equal to or less than about 10 ng, equal to or lessthan about 5 ng, or equal to or less than about 1 ng.

Suitable tissue concentrations of salmeterol via subcutaneousadministration for adipose tissue treatment include from about 1 pM toabout 100 μM, e.g., about 0.01 μM to about 50 μM, 0.5 μM to about 50 μM,about 2.0 μM to about 50 μM, about 5 μM to about 50 μM, about 10 μM toabout 50 μM, about 20 μM to about 75 μM, or any other tissueconcentration of salmeterol from about 0.1 nM to about 100 μM.

Pharmacokinetic Parameters

In another aspect is a pharmaceutical and/or cosmetic formulation,including certain methods of treatment comprising administration of thepharmaceutical and/or cosmetic formulations described herein, comprisingan adipose tissue-reducing amount of salmeterol or a salt, opticalisomer, racemate, solvate, or polymorph thereof and at least onesubcutaneously acceptable inactive ingredient, wherein the formulationprovides a mean plasma salmeterol C_(max) equal to or less than about300 pg/mL when administered subcutaneously. In one embodiment, theformulation provides a mean plasma salmeterol C_(max) equal to or lessthan about 270 pg/mL. In one embodiment, the formulation provides a meanplasma salmeterol C_(max) equal to or less than about 250, about 230,about 200, about 190, about 180, about 170, about 160, about 150, about140, about 130, about 120, about 110, about 100, about 90, about 80,about 70, about 60, about 50, about 40, about 30, about 20, about 10,about 3, about 1 pg/mL, or is undetectable using conventionalmethodology. For purposes of this application, “undetectable usingconventional methodology” or “undetectable using current methodology,”means that the concentration is lower than the low limit of quantitation(LLOQ) using the Liquid Chromatography/Mass Spectrometry/MassSpectrometry (LC/MS/MS) method, which is understood in the art to be atype of tandem mass spectrometry for determining C_(max) levels. Instill further or additional embodiments, the formulations describedherein provide a mean plasma salmeterol C_(max) that is betweenundetectable levels and about 300 pg/mL, about 1 pg/mL to about 270pg/mL, about 5 pg/mL to about 250 pg/mL, about 10 pg/mL to about 220pg/mL, about 15 pg/mL to about 200 pg/mL, about 25 pg/mL to about 170pg/mL, about 50 pg/mL to about 150 pg/mL, or about 75 pg/mL to about 100pg/mL.

Partitioning into Adipose Tissue

In a further aspect is a pharmaceutical and/or cosmetic formulationcomprising an adipose tissue-reducing amount of salmeterol or a salt,optical isomer, racemate, solvate, or polymorph thereof and at least onesubcutaneously acceptable inactive ingredient, wherein the formulationprovides a salmeterol plasma C_(max) ratio of subcutaneous tointravenous administration of between about 0.01 to about 0.4 whenadministered subcutaneously (also known as the “salmeterol partition”ratio) For purposes of this application, the ratio of plasma C_(max) ofa long-acting beta-2 adrenergic receptor agonist administeredsubcutaneously to the plasma C_(max) of the same long-acting beta-2adrenergic receptor agonist administered intravenously is known as the“partition” ratio. Thus, in one embodiment, the partition ratio ofsalmeterol is about 0.01 to about 0.4. In another embodiment, thesalmeterol partition ratio is about 0.05 to about 0.3. In anotherembodiment, the salmeterol partition ratio is from about 0.1 to about0.35. In a further embodiment, the salmeterol partition ratio is about0.1. In another embodiment, the salmeterol partition ratio is between0.05 to about 0.2. In a further embodiment, the salmeterol partitionratio is between about 0.1 to about 0.2. In yet another embodiment, thesalmeterol partition ratio is about 0.01, about 0.02, about 0.03, about0.04, about 0.05, about 0.06, about 0.07, about 0.08, about 0.09, about0.1, about 0.11, about 0.12, about 0.13, about 0.14, about 0.015, about0.16, about 0.17, about 0.18, about 0.19, about 0.2, about 0.21, about0.22, about 0.23, about 0.24, about 0.25, about 0.26, about 0.27, about0.28, about 0.29, about 0.30, about 0.31, about 0.32, about 0.33, about0.34, about 0.35, about 0.36, about 0.37, about 0.38, about 0.39, about0.40. By way of a non-limiting example only, salmeterol xinafoateformulated in 5% PEG-400 in 0.9% saline USP, at various concentrationswere administered to non-naive Gottingen minipigs via single intravenousinjection or subcutaneous injection. The salmeterol partition ratio wascalculated as the average salmeterol plasma C_(max) of subcutaneousadministration ((403+575)/2) divided by the average salmeterol plasmaC_(max) of intravenous administration ((4950+4290)/2). Thus, in thisnon-limiting example, the salmeterol partition ratio was determined tobe 0.1. In a further embodiment, is a subcutaneous formulationcomprising of salmeterol or a salmeterol-like compound and asubcutaneously acceptable excipient wherein the formulation provides apartition ratio of between about 0.01 and about 0.4. As used herein, asalmeterol-like compound is a compound having a partition ratio ofbetween about 0.01 and 0.4 and provides limited systemic exposure, andconsequently, a reduced or minimized risk of producing cardiovascularside effects. Additionally, salmeterol-like compounds also selectivelypartition into the adipose tissue due to their lipophilic nature. Inanother embodiment, provided is a pharmaceutical and/or cosmeticformulation providing a salmeterol partition ratio of between about 0.01to about 0.2. In one embodiment, the formulation provides a salmeterolpartition ratio of between about 0.01 to about 0.3. In yet a furtherembodiment, the formulation provides a salmeterol partition ratio ofbetween about 0.01, about 0.02, about 0.03, about 0.04, about 0.05,about 0.06, about 0.07, about 0.08, about 0.09, about 0.1, about 0.11,about 0.12, about 0.13, about 0.14, about 0.015, about 0.16, about 0.17,about 0.18, about 0.19, about 0.2.

In some embodiments, provided herein is a formulation that is formulatedto provide a single session dose of salmeterol xinafoate in an amountthat is about 5 ng to about 20 μg and that provides a salmeterol plasmaC_(max) ratio of subcutaneous to intravenous administration of betweenabout 0.01 to about 0.4 when administered subcutaneously. In someembodiments, provided herein are pharmaceutical and/or cosmeticformulations comprising a weekly dose of salmeterol xinafoate that isbetween about 5 ng to about 150 μg. In a particular embodiment, providedherein are pharmaceutical and/or cosmetic formulations comprising aweekly dose of salmeterol xinafoate that is less than about 5 μg. Instill further embodiments, provided herein is a pharmaceutical and/orcosmetic formulation comprising a sub-dose of salmeterol xinafoate in anamount that is between about 1 ng to about 50 μg

In a further aspect is a subcutaneous formulation consisting essentiallyof a long-acting beta-2 receptor agonist and a subcutaneously acceptableexcipient thereof, wherein the formulation provides a partition ratiolower than the partition ratio of a reference long-acting beta-2receptor agonist. In one embodiment, the subcutaneous formulationprovides a partition ratio of about four to six times lower than thepartition ratio of a reference long-acting beta-2 receptor agonist. Inone embodiment, the formulation described herein provides a partitionratio about five times lower than the partition ratio of a referencelong-acting beta-2 receptor agonist. In another embodiment, thereference long-acting beta-2 receptor agonist is lipophilic. In yetanother embodiment, the reference long-acting beta-2 receptor agonist isformoterol.

In yet a further embodiment, the salmeterol formulation provides apartition ratio about five times lower than a reference long-actingbeta-2 receptor agonist, wherein the reference long-acting beta-2adrenergic receptor agonist is formoterol.

Methods

Another feature of the subject matter provided herein are pharmaceuticalmethods and cosmetic methods utilizing the formulations provided herein.In some embodiments, provided herein is a cosmetic method for reducingadiposity in a human patient comprising subcutaneously administering apharmaceutical or cosmetic formulation that is suitable for subcutaneousinjection consisting essentially of: (a) active ingredient that consistsessentially of a cosmetically effective adipose tissue-reducing amountof one or more lipophilic long-acting selective beta-2 adrenergicreceptor agonists, or salts, solvates, or polymorphs thereof; and (b)one or more subcutaneously acceptable inactive ingredients. In anotherembodiment, provided herein is a cosmetic method for reducing adiposityin a human patient comprising subcutaneously administering apharmaceutical formulation suitable for subcutaneous injectionconsisting of (a) active ingredient that consists essentially of acosmetically effective adipose tissue-reducing amount of one or morelipophilic long-acting selective beta-2 adrenergic receptor agonists, orsalts, solvates, or polymorphs thereof; and (b) one or moresubcutaneously acceptable inactive ingredients. In still furtherembodiments, provided herein is a cosmetic method for reducing adiposityin a human patient comprising subcutaneously administering apharmaceutical formulation suitable for subcutaneous injectioncomprising: (a) active ingredient that consists essentially of acosmetically effective adipose tissue-reducing amount of one or morelipophilic long-acting selective beta-2 adrenergic receptor agonists, orsalts, solvates, or polymorphs thereof; and (b) one or moresubcutaneously acceptable inactive ingredients. In still furtherembodiments, provided herein is a cosmetic method for inducing lipolysisin adipose tissue of a human patient comprising subcutaneouslyadministering a pharmaceutical formulation suitable for subcutaneousinjection consisting essentially of: (a) active ingredient that consistsessentially of a cosmetically effective adipose tissue-reducing amountof one or more lipophilic long-acting selective beta-2 adrenergicreceptor agonists, or salts, solvates, or polymorphs thereof; and (b)one or more subcutaneously acceptable inactive ingredients. In specificembodiments of the methods and formulations described herein, theinjectable formulations are substantially free of glucocorticosteroids.

Methods of Reducing Adipose Tissue

In another aspect, provided herein are methods for reducing adiposetissue in a patient comprising administering to the patient apharmaceutical and/or cosmetic formulation comprising a lipophiliclong-acting selective beta-2 adrenergic receptor agonist (as singleagent therapy) and at least one subcutaneously acceptable inactiveingredient. In one aspect is a method for reducing adipose tissue in asubject comprising subcutaneously administering to the subject apharmaceutical and/or cosmetic formulation comprising an active agentconsisting essentially of an adipose tissue-reducing amount of alipophilic long-acting selective beta-2 adrenergic receptor agonist or asalt, optical isomer, racemate, solvate, or polymorph thereof and atleast one subcutaneously acceptable inactive ingredient wherein adiposetissue in the subject is reduced. In one embodiment, the lipophiliclong-acting selective beta-2 adrenergic receptor agonist is salmeterol.In yet another embodiment, the salt of the lipophilic long-actingselective beta-2 adrenergic agonist is a xinafoate salt.

In yet a further embodiment, the subcutaneous administration results ina reduced or minimized risk of producing an increase in heart rate or adecrease in blood pressure or a combination thereof. In anotherembodiment, the subcutaneous administration provides a minimized risk ofproducing anaphylaxis related effects, such as, flushing, reddening,rapid heart rate, chest tightness, difficulty breathing, faintness,heart palpitations, hives, atrophy, pigmentation, nodularity, necrosis,irregular or abnormal heart rate, paroxysmal bronchoconstriction, andhypersensitivity reaction such as angioedema and urticaria. By way of anon-limiting example only, a subcutaneous formulation comprisingsalmeterol was administered to Gottingen minipigs. The salmeterolpartition ratio is determined as described above. Thus, in thisnon-limiting example, the subcutaneous formulation administered tominipigs provides a salmeterol partition ratio of 0.1 and a reduced orminimized risk of producing cardiovascular side effects. It should benoted that the reduced or minimized risk described herein (due tolimited systemic exposure) refers to a generalized population and mayvary depending on the individual subject or patient. Subcutaneousformulations consisting essentially of salmeterol provide therapeuticeffect to a regional fat deposit and a reduced or minimized risk ofproducing the side effects associated with the use of other long-actingbeta-2 agonists or long-acting beta-2 agonists administered by othermethods. Such side effects include paradoxical bronchospasm, high bloodpressure, abnormal heart rhythm, abnormally low blood pressure, anincrease in asthma related conditions, bronchospasm, inflammation of thelining of the stomach and intestines, involuntary quivering, fastheartbeat, chest pain, and giant hives.

Methods of Treating Regional Fat Accumulation

In yet another aspect, provided herein are methods for treating regionalfat accumulation in a patient comprising administering to the patient apharmaceutical and/or cosmetic formulation comprising a lipophiliclong-acting selective beta-2 adrenergic receptor agonist (as singleagent therapy) and at least one subcutaneously acceptable inactiveingredient. In further embodiments, the method for treating regional fataccumulation in a subject comprises subcutaneously administering to aregional fat accumulation area a pharmaceutical and/or cosmeticformulation comprising a regional fat accumulation reducing amount ofsalmeterol or a salt, optical isomer, racemate, solvate, or polymorphthereof and at least one subcutaneously acceptable inactive ingredient.In further or additional embodiments, the formulation provides a meanplasma salmeterol C_(max) equal that is in the range of about 300 pg/mLto levels that are undetectable using conventional methodology, whereinthe regional fat accumulation in the subject is reduced. In oneembodiment, salmeterol selectively partitions into adipose tissue of theregional fat accumulation relative to plasma.

Also described herein is a method for treating regional fat accumulationcomprising subcutaneously administering to the subject a pharmaceuticaland/or cosmetic formulation comprising an active agent consistingessentially of a regional fat accumulation reducing amount of along-acting selective beta-2 adrenergic receptor agonist or a salt,optical isomer, racemate, solvate, or polymorph thereof and at least onesubcutaneously acceptable inactive ingredient. In one embodiment, thelipophilic long-acting selective beta-2 adrenergic receptor agonist issalmeterol. In another embodiment, the salt of the lipophiliclong-acting selective beta-2 adrenergic agonist is a xinafoate salt. Ina further embodiment, the subcutaneous administration reduces orminimizes the risk of producing cardiovascular side effects (byminimizing systemic exposure).

Methods of Inducing Lipolysis in Adipose Tissue

As discussed herein, lipolysis and/or inhibition of adipogenesis arestimulated by the beta-1, 2, or 3 receptor subtypes. Thus, agonists toone, two and/or all three receptors are capable of stimulating lipolysisand/or inhibition of adipogenesis. In humans, beta-2 receptor activityis believed to be more important for stimulating lipolysis. Lipolyticactivity and adipocyte proliferation inhibition are believed to bemediated through modulation of adrenergic receptors in adipose tissueand/or on adipocytes.

In another aspect, provided herein is a method for inducing lipolysis inadipose tissue comprising subcutaneously administering a pharmaceuticaland/or cosmetic formulation suitable for subcutaneous injectioncomprising: (a) a lipophilic long-acting selective beta-2 adrenergicreceptor agonist; and (b) at least one subcutaneously acceptableinactive ingredient. In one aspect is a method for inducing lipolysis ina subject comprising subcutaneously administering to the subject apharmaceutical and/or cosmetic formulation comprising an active agentconsisting essentially of an adipose tissue-reducing amount of alipophilic long-acting selective beta-2 adrenergic receptor agonist or asalt, optical isomer, racemate, solvate, or polymorph thereof and atleast one subcutaneously acceptable inactive ingredient that inducedlipolysis in a patient, and reduces adipose tissue in the area treated.In one embodiment, the lipophilic long-acting selective beta-2adrenergic receptor agonist is salmeterol. In yet another embodiment,the salt of the lipophilic long-acting selective beta-2 adrenergicagonist is a xinafoate salt.

Methods of Aesthetic Treatment of Contour Defects Such as Bulging

In an aspect, provided herein is a method for the aesthetic treatment ofbody contour defects or deformities in a human patient comprisingsubcutaneous or transcutaneous administration of a formulationcomprising an adipose tissue-reducing amount of one or more beta-2adrenergic receptor agonists, or salts, solvates, or polymorphs thereof.In certain embodiments, the body contour defect treated is an abdominalcontour defect. In certain embodiments, the body contour defect treatedcomprises abdominal bulging. In some embodiments, the body contourdefect treated is a gluteal contour defect. In an embodiment, the bodycontour defect treated is bulging in an area which is one or more of theneck, upper arms, female and male breasts, abdomen, flanks, back, hips,buttocks, thighs, knees and ankles. In certain embodiments, theindividual being treated has a history of prior treatment of contourdefects (e.g., abdominoplasty, liposuction, or exposure to ablative orbody contouring devices, mesotherapy or lipolytic agents).

In some embodiments, the beta-2 adrenergic receptor agonist administeredfor treatment of contour defects, is selective for the beta-2 adrenergicreceptor. In further or additional embodiments, the beta-2 adrenergicreceptor agonist is lipophilic. In further or additional embodiments,the beta-2 adrenergic receptor agonist is long-acting. In an embodiment,the lipophilic long-acting selective beta-2 adrenergic receptor agonistis salmeterol. In a further embodiment, the polymorph is polymorph I orII. In further embodiments, the formulation uses a mixture of salmeterolpolymorphs. In yet another embodiment, the salt of the lipophiliclong-acting selective beta-2 adrenergic receptor agonist is a xinafoatesalt. In some embodiments, the lipophilic long-acting selective beta-2adrenergic receptor agonist is salmeterol xinafoate.

In a further aspect, provided herein is a method for the aesthetictreatment of body contour defects such as bulging in a human patientcomprising subcutaneously administering a formulation suitable forsubcutaneous injection comprising: (a) active ingredient that consistsessentially of a cosmetically effective adipose tissue-reducing amountof one or more lipophilic long-acting selective beta-2 adrenergicreceptor agonists, or salts, solvates, or polymorphs thereof; and (b)one or more subcutaneously acceptable inactive ingredients. In certainembodiments, the body contour defect treated is abdominal bulging. Insome embodiments, the body contour defect treated is a gluteal contourdefect comprising bulging. In one embodiment, the lipophilic long-actingselective beta-2 adrenergic receptor agonist is salmeterol. In yetanother embodiment, the salt of the lipophilic long-acting selectivebeta-2 adrenergic agonist is a xinafoate salt. In a further embodiment,the subcutaneous administration reduces or minimizes the risk ofproducing cardiovascular side effects (by minimizing systemic exposure).In an embodiment, the, formulation provides a mean plasma salmeterolC_(max) equal that is in the range of about 300 pg/mL to levels that areundetectable using conventional methodology, wherein the body contourdefect in the subject is treated.

In an aspect provided is a method for aesthetic treatment of cheekcontour defect in a human patient by contacting a targeted fat depositin the cheek with a composition comprising an adipose tissue-reducingamount of one or more beta-2 adrenergic receptor agonists, or salts,solvates, or polymorphs thereof. In certain embodiments, the fat deposittargeted is buccal fat. In some embodiments, the fat deposit issubcutaneous cheek fat. In certain embodiments, the patient suffers fromchipmunk cheeks. In some embodiments, the beta-2 adrenergic receptoragonist administered is selective for the beta-2 adrenergic receptor. Infurther or additional embodiments, the beta-2 adrenergic receptoragonist is lipophilic. In further or additional embodiments, the beta-2adrenergic receptor agonist is long-acting. In an embodiment, thelipophilic long-acting selective beta-2 adrenergic receptor agonist issalmeterol. In a further embodiment, the polymorph is polymorph I or II.In further embodiments, the formulation uses a mixture of salmeterolpolymorphs. In yet another embodiment, the salt of the lipophiliclong-acting selective beta-2 adrenergic receptor agonist is a xinafoatesalt. In some embodiments, the lipophilic long-acting selective beta-2adrenergic receptor agonist is salmeterol xinafoate.

In a further aspect provided is a method for aesthetic treatment ofcheek contour defect in a human patient by contacting a targeted fatdeposit in the cheek with a composition comprising: (a) activeingredient that consists essentially of a cosmetically effective adiposetissue-reducing amount of one or more lipophilic long-acting selectivebeta-2 adrenergic receptor agonists, or salts, solvates, or polymorphsthereof; and (b) one or more subcutaneously acceptable inactiveingredients. In an embodiment is a method for aesthetic treatment ofcheek contour defect in a human patient by contacting a targeted fatdeposit in the cheek with a composition essentially comprising: (a)active ingredient that consists essentially of a cosmetically effectiveadipose tissue-reducing amount of one or more lipophilic long-actingselective beta-2 adrenergic receptor agonists, or salts, solvates, orpolymorphs thereof; and (b) one or more subcutaneously acceptableinactive ingredients. In a further embodiment is a method for aesthetictreatment of cheek contour defect in a human patient by contacting atargeted fat deposit in the cheek with a composition consisting of: (a)active ingredient that consists essentially of a cosmetically effectiveadipose tissue-reducing amount of one or more lipophilic long-actingselective beta-2 adrenergic receptor agonists, or salts, solvates, orpolymorphs thereof; and (b) one or more subcutaneously acceptableinactive ingredients. In some embodiments, the patient suffers fromchipmunk cheeks. In certain embodiments, the fat deposit targeted isbuccal fat. In some embodiments, the fat deposit is subcutaneous cheekfat. In one embodiment, the lipophilic long-acting selective beta-2adrenergic receptor agonist is salmeterol. In yet another embodiment,the salt of the lipophilic long-acting selective beta-2 adrenergicagonist is a xinafoate salt. In a further embodiment, the subcutaneousadministration reduces or minimizes the risk of producing cardiovascularside effects (by minimizing systemic exposure). In an embodiment, the,formulation provides a mean plasma salmeterol C_(max) equal that is inthe range of about 300 pg/mL to levels that are undetectable usingconventional methodology, wherein the body contour defect in the subjectis treated.

Methods of Treatment of Thyroid Eye Disease

In an aspect provided herein are compositions, formulations, methods,and systems for treating thyroid eye disease by contacting a targetedfat deposit in the eye with a composition, said composition comprising:one or more beta-2 adrenergic receptor agonists, or salts, solvates, orpolymorphs thereof. In some embodiments is provided a method of treatingproptosis by subcutaneous administration of a composition comprising oneor more lipophilic long-acting selective beta-2 adrenergic receptoragonists, or salts, solvates, or polymorphs thereof

In some embodiments, the beta-2 adrenergic receptor agonist administeredis selective for the beta-2 adrenergic receptor. In further oradditional embodiments, the beta-2 adrenergic receptor agonist islipophilic. In further or additional embodiments, the beta-2 adrenergicreceptor agonist is long-acting. In an embodiment, the lipophiliclong-acting selective beta-2 adrenergic receptor agonist is salmeterol.In a further embodiment, the polymorph is polymorph I or II. In furtherembodiments, the formulation uses a mixture of salmeterol polymorphs. Inyet another embodiment, the salt of the lipophilic long-acting selectivebeta-2 adrenergic receptor agonist is a xinafoate salt. In someembodiments, the lipophilic long-acting selective beta-2 adrenergicreceptor agonist is salmeterol xinafoate.

In a further aspect, provided herein is a method for treating thyroideye disease by contacting a targeted fat deposit in the eye with acomposition comprising: (a) active ingredient that consists essentiallyof an adipose tissue-reducing amount of one or more lipophiliclong-acting selective beta-2 adrenergic receptor agonists, or salts,solvates, or polymorphs thereof; and (b) one or more subcutaneouslyacceptable inactive ingredients. In some embodiments is provided amethod of treating proptosis by subcutaneous administration of acomposition comprising: (a) active ingredient that consists essentiallyof an adipose tissue-reducing amount of one or more lipophiliclong-acting selective beta-2 adrenergic receptor agonists, or salts,solvates, or polymorphs thereof; and (b) one or more subcutaneouslyacceptable inactive ingredients. In some embodiments, the lipophiliclong-acting selective beta-2 adrenergic receptor agonist is salmeterolxinafoate.

Pharmaceutically and Cosmetically Acceptable Excipients

In some embodiments, the pharmaceutical and/or cosmetic formulationsprovided herein comprise a selective, lipophilic, long-acting beta-2adrenergic agonist that is soluble at the desired concentration andtherefore no co-solvent is needed to formulate an injectable solution.

In a further embodiment, the formulations provided herein comprise atleast one co-solvent is selected from about 0.25 to about 40%polyethylene glycol, or from about 0.5 to about 20% polyethylene glycol,or from about 0.75 to about 10% polyethylene glycol, or from about 1 toabout 5% polyethylene glycol, or from about 2 to about 4% polyethyleneglycol. In further or additional embodiments, the polyethylene glycol isabout 0.8-about 1%, or is about 0.9% polyethylene glycol. In a furtherembodiment, the polyethylene glycol is PEG 400.

In yet a further embodiment, the formulations described herein compriseat least one subcutaneously acceptable excipient that is selected fromabout 0.01 to about 10% polysorbate, about 0.05 to about 5% polysorbate,about 0.1 to about 2% polysorbate, or about 0.5 to about 1% polysorbate.In some embodiments, the polysorbate is about 0.01-about 2% polysorbate.In still further embodiments, the polysorbate is about 0.04%. In oneembodiment, the polysorbate is polysorbate 80.

Excipients used in the formulations described herein include, but arenot limited to, suspending agents, surfactants, solubilizers such as,for example, PEG 400, stabilizers, diluents and the like, and should beselected on the basis of compatibility with the lipophilic long-actingbeta-2 adrenergic receptor agonist. In some embodiments, a polyalkyleneglycol or a mixture of different polyalkylene glycols is used as asolubilizer. Polyalkylene glycols from the group of polypropyleneglycols or polyethylene glycols are particularly suitable in thisconnection because of the physiological tolerance. In this connection,the use of polyethylene glycols is utilized in some embodimentspresented herein. In some embodiments, polyethylene glycols such as, forexample, PEG 400, is contemplated herein.

Additives increasing the bioavailability of a lipophilic long-actingbeta-2 adrenergic receptor agonist, such as, salmeterol are, in someembodiments, organic compounds, salts thereof, optical isomers orracemates thereof, emulsions or dispersions containing organic compoundsor salts thereof, e.g. dispersions of polar lipids, or any combination.Organic compounds useful in the subcutaneous formulation are e.g. aminoacids, peptides, proteins, and polysaccharides. Peptides includedipeptides, tripeptides, oligopeptides, such as collagen and gelatin. Insome embodiments, the collagen and gelatin is hydrolyzed.Polysaccharides include e.g., chitosans, cyclodextrins, starch,hyaluronic acids, dextrans, cellulose, and any derivatives,combinations. In further embodiments, the starch is hydrolyzed. Theemulsions include oil-in-water emulsions with oil as the dispersed phaseand water-in-oil dispersions with oil as the continuous phase. In otherembodiments, the oil is of vegetable or of animal origin orsynthetically produced. In further embodiments, the polar liquids areone or more phospholipids or glycolipids or any combination thereof. Insome other embodiments, the additives increasing the bioavailability ofa lipophilic long-acting beta-2 adrenergic receptor agonist, such as,salmeterol are added to a stable solution or dispersion containing thelipophilic long-acting beta-2 adrenergic receptor agonist.

In further embodiments, before administration, one or more aqueoussolutions or dispersions are added, in any mixture or sequence, to thelipophilic long-acting beta-2 adrenergic receptor agonist, such as,salmeterol, which is a stable aqueous solution. In other embodiments,the formulation is a stable aqueous solution ready for administration.In some embodiments, it is a dispersion, e.g. a suspension, a liposomalformulation or an emulsion. In yet other embodiments, the formulationalso comprises a salt in order to give an isotonic solution, e.g., NaCl,KCl, and/or in further embodiments, it comprises one or more otherisotonicity establishing compounds.

In yet other embodiments, an amino acid is used to buffer the system. Insome embodiments, a suitable buffer is glycine, lysine, arginine,histidine or glycylglycine, in other embodiments, the buffer isglycylglycine.

In some other embodiments, a non-ionic surfactant is also present in theformulation. In some embodiments, the surfactant is chosen fromblock-copolymers, such as a poloxamer, e.g., poloxamer 188, or apolyoxyethylene sorbitan fatty acid ester, such aspolyoxyethylene-(20)-sorbitan monolaurate orpolyoxyethylene-(20)-sorbitan monooleate. Also disclosed herein areformulations using polyoxyethylene-(20)-sorbitan monolaurate (Tween 20).In one embodiment, the formulation described herein usedpolyoxyethylene-(20)-sorbitan monooleate (Tween 80). In otherembodiments, the non-ionic surfactant, is present in an amount above thecritical micelle concentration (CMC).

Also presented herein are mono- or disaccharides (e.g., sucrose),polysaccharides such as low molecular weight dextrins, or sugar alcohols(e.g., sorbitol, glycerol or mannitol) used in the subcutaneousformulations. In some embodiments, the formulation also comprisesantioxidants such as bisulfate, ascorbate glutathione, acetylcystein,tocopherol, methionin, EDTA, citric acid, butyl hydroxy toluene and/orbutyl hydroxy anisole. In other embodiments, complexing agents, such asEDTA and citric acid are also present in small concentrations forstabilizing the lipophilic long-acting beta-2 adrenergic receptoragonist, such as, salmeterol. Furthermore, in other embodiments,preservatives such as benzyl alcohol, phenol, sorbic acid, parabens, andchlorocresol are added. In further or additional embodiments, thelipophilic selective long-acting beta-2 adrenergic receptor agonist,such as, salmeterol or formoterol, is prepared as a lyophile. In furtheror additional embodiments, a lyophilized lipophilic selectivelong-acting beta-2 adrenergic receptor agonist is prepared such that itcan be reconstituted for administration via subcutaneous injection to apatient.

Routes of Administration

Injectable formulations are administered using any method known in theart, for example, using a single needle, multiple needles, and/or usinga needleless injection device. In some embodiments, a tissue loadingdose of the active ingredients formulated in a suitable carrierdelivered by injection. In some embodiments, delivery comprises singleneedle injection. In some embodiments, delivery comprises injectionusing a multi-needle array, which, in some embodiments, provides a widedispersion of the formulation in the target tissue. In some embodiments,formulations are injected in a manner that allows dispersal into theappropriate layer of subcutaneous fat in or near regional fat areas.

Transcutaneous formulations, also contemplated as a route of deliveryfor the pharmaceutical and/or cosmetic formulations and methods oftreatment provided herein, are administered using any known method inthe art.

Embodiments of the composition are formulated for administered by anysuitable method, for example, as described in Remington: The Science AndPractice Of Pharmacy (21st ed., Lippincott Williams & Wilkins).Exemplary routes of administration include, but are not limited toparenteral, subcutaneous, or intramuscular. In some embodiments, thecomposition is formulated for injection of an area at which treatment isdesired, for example, in or near a regional fat deposit.

Any suitable pharmaceutically and/or cosmetically acceptable excipientappropriate for a particular route of administration can be used.Examples of pharmaceutically and/or cosmetically acceptable carriersinclude, but are not limited to, buffers, saline, or other aqueousmedia. The compounds described herein are in some embodiments, solublein the carrier which is employed for their administration (e.g.,subcutaneous). Some embodiments comprise any suitable lipophiliccarrier, for example, modified oils (e.g., Cremophor® BASF, Germany),soybean oil, propylene glycol, polyethylene glycol, derivatizedpolyethers, combinations thereof, and the like. Some embodimentscomprise one or more carriers or agents, suitable for subcutaneousadministration. Some embodiments comprise excipients suitable for stablesuspensions for beta-2 receptor agonists. In some embodiments, thepharmaceutical and/or cosmetic formulations comprise polyethylene glycolin an amount of from about 0.5% to about 40%. In another embodiment, theformulation suitable for subcutaneous administration comprisespolyethylene glycol in an amount from about 0.5% to 40%, about 1% toabout 35%, about 2% to about 30%, about 3% to about 25%, about 4% toabout 20%, about 5% to about 15%, about 10% to about 12%, or is about1%, about 2%, about 3%, about 4%, about 5%, about 10%, about 15%, about20%, about 25%, about 30%, about 35%, or about 40%. In a furtherembodiment, polyethylene glycol is in an amount of about 20%. In yet afurther embodiment, polyethylene glycol is PEG 400. In yet anotherembodiment, the pharmaceutical and/or cosmetic formulations comprisepolysorbate in an amount of from 0.01% to about 10%. In anotherembodiment, the formulation suitable for subcutaneous administrationcomprises polysorbate in an amount from about 0.01% to about 10%, about0.02% to about 9%, about 0.03% to about 8%, about 0.04% to about 7%,about 0.05% to about 6%, about 0.06% to about 5%, about 0.07% to about4%, about 0.08% to about 3%, about 0.09% to about 2%, 0.1% to about 1%,about 0.2% to about 0.5%, or is about 0.01%, about 0.02%, about 0.03%,about 0.04%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about0.4%, about 0.5%, about, about 0.6%, about 0.7%, about 0.8%, about 0.9%,about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%,about 8%, and about 9%. In a further embodiment, polysorbate is in anamount of about 10%. In yet a further embodiment, polysorbate ispolysorbate 80. In further or additional embodiments, the lipophilic,selective long-acting beta-2 adrenergic receptor agonist, such as,salmeterol or formoterol, is prepared as a lyophile. In further oradditional embodiments, a lyophilized lipophilic, selective long-actingbeta-2 adrenergic receptor agonist is reconstituted for administrationvia subcutaneous injection to a patient.

In other embodiments, another delivery mode comprises a needlesspressurized injection device. In some embodiments, of these devices, theformulation is pressurized mechanically or pneumatically, for example,using a gas such as helium or carbon dioxide, and then forced through asmall orifice into the body tissues, thereby delivering the formulationsubcutaneously. Suitable formulations for needless injections are known,for example, liquid, solutions, suspensions, gels, colloids, emulsions,and dry powders. An advantage of this system is a wide dispersal areacompared with typical needle injection systems. Needless injection underthe appropriate pressure forces the formulation into a more planardelivery pattern, with fingers of formulation spreading out radiallyfollowing paths of least resistance. In contrast, delivery by a typicalneedle injection results in a globular delivery of the formulation.Needless injection also permits precise control of the depth ofpenetration by controlling the injection pressure and orifice size.Thus, needless injection is a delivery method for a sub-dermal injectioncontemplated herein of a formulation for treating superficial fataccumulations, which is useful, for example, for smoothing skin dimplingcaused by fat. In other embodiments, needless injection is also used fordeeper fat accumulations. In further embodiments, a needless device alsoprovides easy and convenient multiple injections of the formulation overa defined region with a large lateral spread.

Treatment of Other Conditions

In some embodiments, the formulations described herein are used fortreating immune and inflammation-related dermal conditions includingpsoriasis, atopic dermatitis, vitiligo, hypopigmentation, stria, andwrinkles or rhytids. In one embodiment, a lipophilic, selectivelong-acting selective beta-2 agonists is administered subcutaneously totreat a condition of a human patient. In another embodiment, a betaagonist is used for treating immune and inflammation-related dermalconditions. In some embodiments, subcutaneously administrablelipophilic, selective long-acting beta-adrenergic agonist treatment isused to decrease Langerhans cell migration and antigen presentation.

In some embodiments, beta-2 adrenergic receptor agonists areadministered subcutaneously for the treatment of skin wrinkles and skinstria, or stretch marks. Cutaneous stria is characterized by a thinningof the dermis, with a loss of collagen and hypopigmentation. Long-actingbeta-adrenergic agonists promote the recruitment and proliferation andcollagen production of dermal fibroblasts in the stria. In addition,they stimulate melanocytes to repigment the stria. Thus, in someembodiments a subcutaneous formulation consisting essentially of anadipose tissue-reducing amount of salmeterol or a salt, solvate, orpolymorph thereof is used to treat these conditions.

In some embodiments, beta-2 adrenergic receptor agonists areadministered to increase skeletal muscle mass and cause hypertrophy andincreased protein synthesis, effects which are mediated throughintracellular in increases cAMP levels. Similar to adipocytes, exposureto beta-2 adrenergic receptor agonists results in receptordown-regulation. Thus, the disclosed formulations are also used fortreating skeletal muscle injury or conditions where increasing skeletalmuscle mass is important. In some embodiments, the methods describedherein are used to increase facial muscle tone and provide a moreyouthful appearance. In some embodiments, the methods described hereinare used to treat strabismus or lazy eye by strengthening ocularmuscles.

In other embodiments, provided are methods for decreasing cellulite in asubject, also known as adiposis edematosa, dermopanniculosis deformans,status protrusus cutis, and gynoid lipodystrophy, comprisingadministering via subcutaneous methods, a subcutaneous formulationconsisting essentially of a long-acting beta-2 adrenergic receptoragonist and a subcutaneously acceptable excipient thereof, wherein theformulation decreases cellulite in the subject.

Areas of fat deposits on a subject, such as for example a human patient,for which the formulations described herein are useful include, but arenot limited to, the inside region of the knees, the middle to upper areaof the upper arm, including the tricep area, the submental area,including the area under the chin, for example the wattle (which isunderstood to refer to the fleshy fold of skin in the submental area ofa patient), the abdomen, the hips, the inner thigh, the outer thigh, thebuttocks, an upper arms region of the patient, the lower back and thechest.

In some embodiments, inducing lipolysis and inhibiting fat cell growthin regional fat accumulations, have additional health benefits throughthe shrinkage of the average fat cell diameter or volume. Large volumefat cells actively secrete pro-inflammatory and deleterious hormonessuch as TNF-alpha and interleukins (“adipokines”), which contribute tocomorbidities associated with fat, such as diabetes. By reducing thesize of these fat cells and therefore the deleterious adipokinesecretion, improvements in fat-related comorbidities are realized.

In some embodiments, the disclosed formulations (e.g., subcutaneous andtranscutaneous formulations), are used for treating obstructive sleepapnea. Obstructive sleep apnea occurs when the airway is temporarilyblocked during sleep, leading to hypoxia, high blood pressure, cardiacdysrhythmia, and a higher risk of death. Excessive fat in the pharynxand soft palate it believed to contribute to this blockage. Obese peoplehave a higher incidence of sleep disorders and persons with sleep apneahave excessive fat in the palate and pharynx on MRI. Thus, in someembodiments, formulations described are administered to a subject toreduce the symptoms of sleep apnea. In some embodiments, theformulations are administered locally (e.g., by injection) into thepalate or pharynx transorally. In some embodiments, the formulations areadministered by subcutaneous into the region the neck to reduce theobstructive symptoms.

While certain embodiments have been described, these embodiments havebeen presented by way of example only, and are not intended to limit thescope of the disclosure. The formulations, methods, and systemsdescribed herein may be embodied in a variety of other forms.Furthermore, various omissions, substitutions and changes in the form ofthe formulations, methods, and systems described herein may be madewithout departing from the spirit of this disclosure. The accompanyingclaims and their equivalents are intended to cover such forms ormodifications.

Examples

The following specific examples are to be construed as merelyillustrative, and not limitative of the remainder of the disclosure inany way whatsoever. The examples described herein reference and providenon-limiting support to the various embodiments described in thepreceding sections.

Example 1: Plasma Pharmacokinetics Following Single Intravenous andSubcutaneous Injections of a Lipophilic Long-Acting Selective BetaAgonist

Materials and Methods:

Salmeterol xinafoate was formulated in 5% PEG-400 in 0.9% saline, USP,at concentrations of 0.1, 1, 10, and 100 μg/mL. 1 male and 1 femalenon-naive Gottingen minipigs (18.4 to 19.5 kg at the time of the IVdose) were utilized. The animals were housed individually. The dosingscheme is shown in Table 1 below:

Injection # Route Dose (μg/kg) No. Animals 1 IV 4 1 2 SC 4 1 3 SC* 4 1 4SC* 0.004 1 5 SC* 0.04 1 6 SC* 0.4 1 *The dose volume was equallydivided over 5 injection sites

Each minipig received a single IV injection followed by SC doses of thesalmeterol formulation. The IV dose was administered by slow bolus (1minute; 0.0167 hr) into the marginal ear vein, whereas the first SC dosewas administered as a bolus injection along the back (neck-region) wherethere was a longitudinal fat depot. The second SC dose was distributedin 5 separate SC injection sites. The fourth, fifth, and sixthadministrations were SC administrations divided over 5 injection sits ofdifferent graded concentrations (0.1, 1, and 10 μg/mL of salmeterol in5% PEG). Each dose was separated by at least a 3-day washout period.Blood samples (approximately 4 mL) were collected via thebranchiocephalic plexus at pre-dose and at 2.00, 5.00, 10.0, 20.0, 30.0and 45.0 minutes, and at 1.00, 2.00, 4.00, 8.00 and 24.0 hr post-dose.Blood samples were placed in tubes containing K₂-EDTA as theanticoagulant. Samples were processed to plasma by centrifugation andstored frozen at approximately −70° C. (+/−15° C.) until shipment foranalysis.

Sample Analysis:

Plasma samples were analyzed for salmeterol using a qualified liquidchromatography/mass spectrometry/mass mass spectrometry (LC/MS/MS)method. The lower limit of quantitation (LLOQ) was 2.50 pg/mL. Sampleswere analyzed within a maximum of 14.0 days of collection.

Data Analysis:

Non-compartmental pharmacokinetic parameters were calculated usingWinNonlin 5.2 software, NCA model 202, IV infusion input for the IV dataand NCA model 200, extravascular input for the SC data. Individualplasma concentrations for each animal were used in the calculation ofpharmacokinetic parameters. Nominal collection times and doses were usedin the calculations. The area under the plasma concentration-time curve(AUC) was calculated using linear trapezoidal approximation (linear/loginterpolation). Concentration values below the assay limit ofquantitation were set to zero for calculations. The maximum plasmaconcentration (C_(max)) and the time of its occurrence (T_(max)) wereverified by inspection. The half-life (t_(1/2)) values were calculatedusing the last 3 plasma concentrations with nonzero values, if datapermitted.

Results:

The pharmacokinetic parameters are shown in Table 2. The plasmaconcentrations of salmeterol are shown in Table 3. The plasmaconcentrations of salmeterol and formoterol (via subcutaneous andintravenous administration) are shown in FIG. 1. Actual sampling timesdiffered from nominal sampling times on occasion by more than 10%. Datain the apparent elimination phase were inadequate to calculate somepharmacokinetic parameters for some animals (indicated as ND for notdetermined or NA for not applicable). Bioavailability was likely poorlyestimated due to scarcity of data in the apparent terminal eliminationphase to calculate the apparent terminal elimination rate constant. Thisaffected calculation of AUC_(inf), t_(1/2), CL, V_(ss), and F. Thesedepartures are not believed to have significantly impacted the overallpharmacokinetic conclusions.

TABLE 2 Pharmacokinetic Parameters of Salmeterol Following IV and SCAdministration to Gottingen Minipigs T_(max) C_(max) AUC_(inf) t_(1/2)t_(last) CL V_(ss) Route Sex Animal (hr) (Pg/mL) (pg · hr/mL) (hr) (hr)(mL/hr/kg) (mL/kg) IV Female 2-0623 0.0333 4950 1470 9.52 24.0 273022100 IV Male 1-0771 0.0333 4290 1750 7.89 24.0 2280 17100 SC1 4 Female2-0623 1.00 403 1850 6.02 24.0 1.26 SC5 4 Female 2-0623 0.0333 626 20108.58 24.0 1.37 SC1 4 Male 1-0771 0.0833 575 1290 5.92 24.0 0.737 SC5 4Male 1-0771 0.0833 681 1510 6.84 24.0 0.863 SC5 0.004 Female 2-0623 1.003.08 ND ND 1.00 ND SC5 0.004 Male 1-0771 NA 0 ND ND NA ND SC5 0.04Female 2-0623 0.0333 8.31 ND ND 0.0833 ND SC5 0.04 Male 1-0771 0.08332.77 ND ND 0.0833 ND SC5 0.4 Female 2-0623 0.0333 113 164 2.88 8.00 1.12SC5 0.4 Male 1-0771 0.0833 64.5 138 2.94 8.00 0.789 AUC_(inf) = areaunder the curve at infinite time. CL = plasma clearance. F =bioavailability; NA = not applicable; ND = not determined; SC1—singlesite injection; SC5 = SC injection split among 5 sites. t_(last) = timeof last measurable plasma concentration. V_(ss) = volume of distributionat steady state The IV dose was 4 μg/kg The injection 4 SC5 0.004 μg/kgmale showed no plasma concentrations >LLOQ of salmeterol at any timepoint.

TABLE 3 Plasma Concentrations of Salmeterol Following IV and SCAdministration to Gottingen Minipigs (08-529) Dose Time (hr) Inj Rte(mg/kg) Sx 0 0.0333 0.0833 0.167 0.333 0.500 0.750 1.00 2.00 4.00 8.0024.0 1 IV 4 M < 4290 1380 502 326 240 177 161 121 70.6 53.6 12.5 1 IV 4F < 4950 1300 559 322 232 152 154 81.1 49.1 35.3 11.3 2 SC 4 M < 495 575423 284 278 236 212 168 74.5 30.6 6.23 2 SC 4 F < 124 227 206 334 302351 403 206 108 54.4 10.0 3 SC5 4 M < 604 681 519 449 483 278 251 14875.6 36.1 8.92 3 SC5 4 F < 626 485 442 398 435 435 388 196 86.8 51.016.1 4 SC5 0.004 M < < < < < < < < < < < < 4 SC5 0.004 F < < < < < < <3.08 < < < < 5 SC5 0.04 M < < 2.77 < < < < < < < < < 5 SC5 0.04 F < 8.313.17 < < < < < < < < < 6 SC5 0.4 M < 32.0 64.5 57.0 43.3 53.8 26.5 28.719.5 8.46 4.41 < 6 SC5 0.4 F < 113 55.4 42.9 59.8 83.5 34.5 32.2 24.58.58 5.15 < Plasma concentrations are reported in pg/mL. LLOQ = 2.50pg/mL. < = LLOQ; F = female; Inj = Injection number; M = male; Rte =route; SC1 = single site injection; SC5 = SC injection split among 5sites; Sx = sex. The male animal was number 1-0771 and the female animalnumber was number 2-0623

Overall there were no consistent, substantial differences in C_(max) orAUC values between the male and female animal. Additionally, there wasnot a substantial difference in C_(max) and AUC_(inf) values betweensingle site SC doses and multiple (5) site SC doses at the 4 μg/kg dose.The ratios (SC1/SC5) ranged from 0.644 to 0.844 for C_(max) and 0.854and 0.920 for AUC_(inf). Salmeterol appeared to be well absorbed afterSC administration at a single site at 4 μg/kg and after divided SCadministration at 5 sites at the 4 and 0.4 μg/kg doses; bioavailabilityranged from 0.737 to 0.863 for the male and 1.26 to 1.37 for the femalefor the 4 μg/kg dose regardless of the number of sites of administrationand 0.789 for the male and 1.12 for the female for the 0.4 μg/kg dose at5 divided SC sites. Scarce or no plasma concentrations were observed forthe 0.04 and 0.004 divided SC doses.

Conclusion:

In this single male and female Gottingen minipig study, salmeterolappeared to be well absorbed after subcutaneous administration at asingle site and after divided SC administration at 5 sites at the 4μg/kg dose and after divided SC administration of the 0.4 μg/kg dose.Overall there were no consistent, substantial differences in Cmax andAUC_(inf) values between single site SC doses vs. multiple site SCdoses. Overall there were no substantial differences in pharmacokineticparameters between the male and female animals.

Example 2: Human Clinical Trial Comparing Lipophilic Long-ActingSelective Beta-2 Adrenergic Receptor Agonists with Glucocorticosteroids,their Combination, and Placebo

Title A Multi-center, Randomized, Double-Masked, Placebo-controlled,Multi-Factorial Evaluation of the Safety and Efficacy of a monotherapylipophilic long-acting selective beta-2 adrenergic receptor agonist(e.g., salmeterol xinafoate (SX)) compared with a monotherapyglucocorticosteroid (e.g., fluticasone Propionate (FP)), a combinationof FP and SX, and placebo for the Reduction of Subcutaneous AbdominalAdiposity Study Objectives This placebo-controlled factorial studyexamines the comparative efficacy and safety of SX alone, FP alone, andof the combination of SX + FP to reduce abdominal subcutaneous adiposetissue. To evaluate the safety and efficacy of subcutaneous injectionsof 0.02 μg SX/mL once weekly for 8 weeks for the reduction of abdominaladiposity. To evaluate the safety and efficacy of subcutaneousinjections of 0.02 SX/mL once weekly for 8 weeks for the treatment ofabdominal contour defects. To evaluate the safety and efficacy ofsubcutaneous injections of 1.0 μg FP/mL once weekly for 8 weeks for thereduction of abdominal adiposity. To evaluate the safety and efficacy ofsubcutaneous injections of SX + FP combo (1.0 μg FP + 0.02 μg SX/mL onceweekly for 8 weeks) for the reduction of abdominal adiposity. StudyDesign A double-masked, multiple-dose study of the safety and efficacyof 0.02 μg/mL of SX alone compared to 1.0 μg/mL of FP alone, 1.0/0.02μg/mL (FP/SX) combo, and to placebo. Doses will treat a region ofadiposity and/or contour defect and will be administered as twenty 1 mLsubcutaneous injections once a week for 8 weeks in 200 subjects withmeasurable abdominal subcutaneous adipose tissue. The total SX dose of0.4 μg administered in 20 mL in this study is approx. 100 times lowerthan the established salmeterol dose that yields peak serumconcentrations (Cmax) and systemic exposure (AUC) equivalent to thereference-listed product ADVAIR DISKUS ® 500/50. Study Drug SX is asterile, preservative-free, clear, aqueous solution salmeterol xinafoatefor injection. The drug products used in the study will be supplied asSalmeterol Xinafoate for Injection, 10 μg. A lyophile of the SX drugproduct will be supplied in separate 3 mL glass vials to bereconstituted with Sterile Water for Injection. Immediately prior toadministration, the lyophile drug products will be reconstituted anddiluted with Sterile Saline Solution USP (0.9% Sodium Chloride). Thesupplied drug products (Salmeterol Xinafoate for Injection andFluticasone Propionate for Injection) should be stored at roomtemperature (15° C. to 25° C.). The reconstituted solution of each drugproduct (the SX, FP, and combo) should be protected from light untiladministration. The Salmeterol Xinafoate for Injection and FluticasonePropionate for Injection drug products will be provided in bulk nestedpackaging. Placebo: Sterile Saline, USP (0.9% Sodium Chloride) will beused as the placebo. Treatment Groups Eligible subjects in thismultiple-dose study will be randomized into four groups of 50 to receivetwenty 1 mL subcutaneous injections of either a combination of 1.0 μg/mLFP + 0.02 μg/mL SX, or 1.0 μg/mL FP alone, or 0.02 μg/mL SX alone, orPlacebo. The 20 subcutaneous injections will be spaced approximately 4cm apart and will treat a pre-marked abdominal area of adiposity that isapproximately 400 cm². The sequence of once weekly subcutaneousinjections will continue for 8 consecutive weeks for a total of 160injections administered into the marked abdominal area. Comprehensivesafety assessments will be performed at each subject visit through theEnd of Study Visit on Day 78 ± 2 days. Duration of Screening period: upto 30 days. Treatment The expected study duration is 12 weeks comprisedof an 8 week treatment period and 1 and 4 week post-therapy follow-upvisits for assessment of safety and efficacy. Duration Approx. 7 monthsStudy Population The study population will consist of qualified male andfemale subjects, 18 years of age and older, inclusive who have providedwritten, informed consent. Qualified subjects, who meet all inclusionand exclusion criteria, will be enrolled into the study. Total No. of Atleast 200 subjects will be enrolled. The final subject number will bedetermined Subjects following an interim analysis to be conducted once180 subjects have completed treatment or discontinued from the study.Subjects who give informed consent in writing but who do not receivestudy drug for any reason will be considered screen failures and will bereplaced. Sites Up to 10 sites Inclusion Criteria 1) Healthy male andfemale patients, 18 years and older, who have provided written, informedconsent prior to any study procedures (including pre-treatment andscreening) being performed 2) Patients who have a localized area ofabdominal subcutaneous adiposity, which is Grade 3 or higher on thePatient and Clinician Photonumeric Scales 3) Patients who havesatisfactory abdominal skin elasticity as assessed by the investigator4) BMI ≦ 25 kg/m2 and stable body weight as shown by a weight variance ≦3% between Screening and Day 1 5) History of a stable diet and exerciseroutine in the 3 months prior to Screening 6) Female subjects who have anegative urine pregnancy test at Screening and Day 1, and who agree touse adequate birth control methods (abstinence, female partner,stabilized on oral contraceptives for at least two months, implant,injection, IUD, patch, NuvaRing ®, condom and spermicidal, diaphragm andspermicidal) throughout the entire study until completion of the Week 12End of Study Visit procedures Exclusion Criteria 1) History of priortreatment of abdominal subcutaneous adipose tissue (e.g.,abdominoplasty, liposuction, or exposure to ablative or body contouringdevices, mesotherapy or lipolytic agents) 2) Female subjects who arewithin 12 months post-partum, or who are pregnant, lactating, and/or whoare of childbearing potential but not using adequate birth controlmethods 3) Any skin conditions in the treatment area that may affectstudy procedures or evaluations - including but not limited to skininfections, psoriasis, eczema, tattoos, striae, keloids, or hypertrophicor tethered scars, or excessive skin wrinkles, or a pannus 4) Subjectsplanning to embark on a weight loss or exercise program during studyparticipation 5) Subjects who partake of abdominal massaging and who areunwilling to discontinue this therapy during the study 6) Knownhypersensitivity to the study drugs and/or any of their components 7)Prior or current enrollment in any Lithera study involving SX, FP, orSX + FP. 8) Concurrent enrolment in another investigational drug ordevice study; or use of any experimental or investigational drug ordevice within 30 days, or for drugs within 6 times the eliminationhalf-life prior to Day 1 if that is longer 9) Any medical condition thatin the opinion of the investigator might jeopardize the subject's safetyor complicate study procedures or assessments, including, but notlimited to: (a) any bleeding, or connective tissue disorders; (b)asthma, COPD, diabetes (Type I and II) or cardiovascular disease(subjects with well- controlled hypertension will not be excluded unlessthey are taking beta-blocker drugs); (c) history of major surgery within30 days prior to Day 1, or planned surgery during the study period; (d)any lymphatic disease causing lymphedema in the treatment area; (e)abdominal hernias, abdominal recti muscle divarication or diastasis,visible abdominal organomegaly, or abdominal asymmetry due tomusculoskeletal abnormalities; (f) a history of any DSM-IV psychiatricdisorder; (g) any clinically-significant physical examination findings,as determined by the Investigator, at Screening or Day 1; (h) anyclinically-significant abnormal laboratory result during Screeningand/or Day 1, as determined by the investigator 10)  Use of drugs withanticoagulant activity (including aspirin and NSAIDs), immunomodulators(including steroids), anti-metabolites, β-adrenergic receptor agonistsor blockers, strong CYP 3A inhibitors, or nonpotassium -sparingdiuretics (e.g., loop or thiazide diuretics) prior to Day 1 during thestudy 11)  Use of tricyclic antidepressants or monoamine oxidaseinhibitor medications within 14 days prior to Day 1, or during the study12)  Subjects unlikely or unable to comply with protocol procedures oradhere to the study visit schedule Procedures All injections will beperformed in an outpatient setting. At each visit, subjects will receivea total of 20 subcutaneous injections (1 mL) to infiltrate an area ofapproximately 400 cm². Subjects will maintain their usual diet andexercise routine during the study: any fat treatment, including but notlimited to liposuction, mesotherapy and abdominal massaging will not beallowed. Screening Procedures Subjects will undergo screening proceduresat the Screening Visit. This visit must occur within 30 days (Day −30 toDay 0) prior to study randomization at Day 1. Study procedures will beexplained to each subject and written, informed consent must be obtainedprior to initiating any study-related procedures, including screeningprocedures. Qualified subjects, who meet all Inclusion/Exclusioncriteria, with baseline screening laboratory tests results within normallimits as defined per protocol, will be scheduled for the RandomizationVisit on Day 1. It is required that all Randomization Visits (Day 1) arescheduled to ensure that over the 8 week treatment period, weekly studydrug administration for each subject occurs in a regular cycle, withdoses of study drug administered on the same day each week (±2 days). OnStudy Procedures Randomization and Treatment Visit (Day 1) Pre-dose (Day1): Eligible study subjects will visit the clinic on Day 1 and pre-doseprocedures will be performed according to the Schedule of Events. Forsubjects continuing to meet all of the Inclusion/Exclusion criteria,baseline standardized digital photographs (using the Canfield Vectra ™CR-10 16 mm 360° 3-D imaging system). Immediately thereafter, abdominalskin markings for the injection sites will be placed using the treatmentarea grid in order to ensure consistent study drug administration overthe course of the study and a baseline abdominal circumferentialmeasurement using a constant tension tape measure at pre-marked levelsin the treatment area will be taken before dosing. Subjects then will berandomized to double-masked study drug. According to randomization,twenty 1.0 mL subcutaneous injections of either FP + SX, or 1.0 μg/mL FPalone, or 0.02 μg/mL SX alone, or Placebo will be administered into theabdominal treatment area using the injection grid. Post-dose (Day 1):After study drug administration, post-dose procedures will be performedaccording to the Schedule of Events. Subjects must remain in the clinicuntil the post-dose assessments have been completed and during this timeconcomitant medications and adverse events will be reviewed andrecorded. Before leaving the clinic, all subjects will be scheduled fortheir next visit. A subject will not be discharged from the clinic if,in the Investigator's opinion, the subject has an adverse event thatrequires further in-clinic monitoring. Outpatient On-Treatment Visits(Days 8. 15. 22. 29. 36. 43. 50 ± 2): Subjects will receive subcutaneousinjections of study drug administered weekly for 8 weeks. At each studyvisit, 20 injections will be administered subcutaneously using astandard injection grid with the individual 1 mL injections being spacedapproximately 4 cm apart. This schedule will be repeated for eachsubject for a total of 8 weekly treatment sessions during the study.Pre-dose (Days 8. 15. 22. 29. 36. 43. 50 ± 2): At each Treatment Visit,pre-dose procedures will be performed as per the Schedule of Events(TABLE 1). Prior to study drug being injected on the dosing day of eachweek, circumferential tape measurements of the treatment area, at thepre-marked levels in the treatment area will be made. The skin markingson the abdomen will be used to align an injection grid that will be usedto guide the administration of the injections on the assigned dosing dayeach week. The treatment area will be standardized, and will involve 20individual subcutaneous injections at each treatment visit. In addition,on Day 29 ± 2, a standardized measurement of the treatment area using adigital 3-D photographic device will be made prior to the application ofthe temporary injection grid. Post-dose (Days 8, 15, 22, 29, 36, 43, 50± 2): On all treatment visits, the post-dose procedures will beperformed as per the Schedule of Events. Subjects must remain in theclinic until the post-dose assessments have been completed and duringthis time concomitant medications and adverse events will be reviewedand recorded. Before leaving the clinic, subjects will be scheduled fortheir next visit. A subject will not be discharged from the clinic if,in the Investigator's opinion, the subject has experienced an adverseevent that requires further in-clinic monitoring. 1 Week Post-TreatmentVisit (Day 57 ± 2): A week after the last study drug administration onDay 57 ± 2, patients will return to the clinic for their initialpost-treatment visit. The study procedures will be performed as per theSchedule of Events. On Day 57 ± 2 a standardized measurement of thetreatment area using 3-D photography will be performed. Then, abdominalcircumference measurements using a tape measure at the pre-marked levelsin the treatment area will be recorded. Any abnormal laboratory teststhat are clinically significant will be followed until resolution and/orrepeated per protocol. End of Study Visit (Day 78 ± 2) or EarlyTermination Visit: Four weeks after the last study drug administrationon Day 78 ± 2, patients will return to the clinic for their End of Studyvisit. The study procedures will be performed as per the Schedule ofEvents. On Day 78 ± 2 a standardized measurement of the treatment areausing 3-D photography will be performed. Then, abdominal circumferencemeasurements using a tape measure at the pre-marked levels in thetreatment area will be recorded. If a subject withdraws from the studyearly, and the Day 78 ± 2 End of Study Visit procedures have not beenperformed, these procedures should be performed prior to terminating thesubject from the study. Any abnormal laboratory tests that areclinically significant will be followed until resolution and/or repeatedper protocol. Once the End of Study Visit procedures have beenperformed, the subject has completed the study. Safety Assessments Thefollowing safety assessments will be performed at the designated timepoints as specified in the protocol: (1) vital signs (systolic anddiastolic blood pressure, heart rate, breathing rate and bodytemperature); (2) physical examinations; (3) clinical assessment ofinjection site reactions (local tolerability using the Injection SiteReaction Severity Scale); (4) clinical laboratory tests (hematology,serum chemistry, and urine dipstick analysis); (5) adverse events.Safety parameters monitored during this study will be compared withineach treatment group (i.e. changes from baseline) and between treatmentgroups. Efficacy Efficacy assessments include: (1) circumferentialmeasurements derived from analysis of Assessments standardized 3-Dphotographs of the treatment area; (2) manual tape circumferentialmeasurement of the abdominal treatment area at the level of theumbilicus, and at other fixed levels in the treatment area; and, (3)volumetric changes in the treatment area derived from analysis ofstandardized 3-D photographs of the treatment area. Study Endpoints Thestudy endpoints will include both safety assessments and evaluations ofefficacy of the four treatments being assessed. The study populationwill be evaluated for several objective efficacy assessments based onstandard measurements of subcutaneous adipose tissue during this study,including: (1) manual tape circumferential measurement of the abdominaltreatment area at the level of the umbilicus, and at other fixed levelsin the treatment area; (2)circumferential measurements derived fromanalysis of standardized 3- D photographs of the treatment area; and (3)volumetric changes in the treatment area derived from analysis ofstandardized 3-D photographs of the treatment area. Sample Size Calc.Initially 50 subjects per treatment group (total N = 200) withobservable abdominal subcutaneous adipose tissue (per the InclusionCriteria) will be enrolled (and have signed an IC form and received atleast one dose of study drug). As the assumption of variability greatlyaffects the estimate for sample size, an interim review to assess thechange from baseline in abdominal measurements will be conducted toassess the adequacy of the overall sample size relative to achieving thestudy objectives. After approximately 180 subjects have completed thestudy, an interim analysis of the primary efficacy results only will beconducted by an independent statistician. This analysis will be usedonly to confirm the adequacy of the planned sample size or to recommendenlarging the sample size to achieve adequate statistical power todetect the hypothesized difference between the SX, FP, and FP + SXtreatment groups. If the re-estimate of the sample size results in astudy size that is not practical to complete, Lithera will complete thestudy as originally designed (Shun et al., 2001). Statistical AnalysisDescriptive statistical analyses will be presented for all efficacy andsafety endpoints. Summary statistics will include counts and percentagesfor categorical variables and the number of subjects, mean, standarddeviation (SD), median, minimum and maximum for continuous variables.Continuous efficacy endpoints will be summarized by treatment groupusing descriptive statistics. Differences between treatment groups willbe assessed using an ANCOVA with effects for treatment, site, andbaseline. Compliance This protocol was developed according to theguidelines of the International Conference on Statement Harmonisation(ICH) Good Clinical Practice (GCP) Guidelines E6 and 21 Code of Federal

Example 3: Pharmaceutical and Cosmetic Formulations SubcutaneousFormulation

To prepare a parenteral pharmaceutical and/or cosmetic compositionsuitable for subcutaneous administration, a lipophilic, selective,long-acting beta agonist, such as salmeterol xinafoate, is dissolved inPEG 400 which is stabilized with polysorbate 80. Water is then added.This solution is stored in a single-use glass vial which is storedfrozen and protected from light until dose preparation. When ready foruse, the salmeterol solution is then diluted to a suitable concentrationfor subcutaneous administration using a diluent made of an aqueoussolution of 20% PEG 400, 1% polysorbate 80, and sterile water forinjection. Optionally, in certain embodiments, a co-solvent is used. Inother embodiments, a co-solvent is not used. For example, in certainembodiments, a lipophilic, selective, long-acting beta agonist islyophilized.

Transcutaneous Formulation

Also provided herein is a transcutaneous formulation for administrationto a patient, and methods of treatment provided herein, including forexample methods for the reduction of adipose tissue, using thetranscutaneous formulations provided herein. In some embodiments, alipophilic, selective, long-acting beta agonist is formulated in atranscutaneous formulation which comprises about 0.5% to about 10% byweight salmeterol xinafoate or formoterol fumarate, about 1% to about75% propylene glycol or isopropyl alcohol, and optionally otherexcipients including but not limited to transcutol, propyl gallate,water, and ethanol, wherein the total percent by weight is 100%.

Example 4: Clinical Testing for Treatment of Graves' Ophthalmopathy withCompositions Comprising a Lipophilic Long-Acting Selective Beta Agonist

A non-limiting example of such a clinical testing for treatment ofGraves' Ophthalmopathy is as follows:

Patient Selection:

Patients are to be 18 years of age or above and have no hypersensitivityto the administered drugs. They are diagnosed with proptosis symptomsassociated with Graves' Ophthalmopathy by ultrasonography orcomputerized tomography. In particular, patients are chosen withunilateral or bilateral proptosis edema of 3 mm or more, with or withouteyelid swelling. Patients may also exhibit diplopia, limitation of eyemovement in extreme positions, and evident restriction of movement.Patients may have undergone thyroidectomy for hyperthyroidism. Othersteroid therapies should be not used for treatment of hyperthyroidism.All studies are to be performed with institutional ethics committeeapproval and patient consent.

Study Design:

Test 1:

This is a multicenter, dose escalation study of the therapy ofsalmeterol, a long acting beta 2 agonist in the treatment of thyroid eyedisease. Patients receive an injection administration of a parenteralcomposition of the drug daily. Patients who do not achieve proptosisimprovement or partial or complete response but who have stable diseaseafter 1 week of therapy will receive an additional 1 week of therapy ata higher dose than what is originally assigned. Cohorts of 3-6 patientsreceive escalating doses of the drug until the maximum tolerated dose(MTD) is determined. The MTD is defined as the dose preceding that atwhich 2 of 3 or 2 of 6 patients experience dose-limiting toxicity.

Test 2:

This is a randomized, multicenter study. The study length is 60 days.Patients are randomized to 1 of 18 treatment groups. For group 1,patients are given salmeterol alone daily at MTD. For Group 2, patientsare given salmeterol on every other day. For Group 3, patients are givensalmeterol once a week. Groups 4-6 have the same dosing regime as 1-3except the dosage is at one-fourth MTD. Groups 7-9 also have the samedosing regime as 1-3 except the dosage is at one-tenth MTD. In additionto the treatment groups, a control group is left untreated.

Endpoint Assessment:

Patients are assessed for reduction of proptosis and decrease of orbitalfat volume and extraocular muscles at the conclusion of the study.Improvement in eyelid closure and ocular movement is also assessed. A20% reduction within 60 days is presumed as a positive outcome.

Example 5: Testing for Aesthetic Treatment of Cheek Contour Defects withCompositions Comprising a Lipophilic Long-Acting Selective Beta Agonist

A non-limiting example of such a clinical testing for aesthetictreatment of cheek contour defects is as follows:

Patient Selection:

Patients are to be 18 years of age or above and have no hypersensitivityto the administered drugs. Other steroid therapies should be not used.All studies are to be performed with institutional ethics committeeapproval and patient consent.

Study Design:

Test 1:

This is a multicenter, dose escalation study of the therapy ofsalmeterol, a long acting beta 2 agonist in the aesthetic treatment ofcheek contour defects. Patients receive an injection administration of aparenteral composition of the drug daily. Patients who do not achievereduction of buccal or subcutaneous cheek fat after 1 week of therapywill receive an additional 1 week of therapy at a higher dose than whatis originally assigned. Cohorts of 3-6 patients receive escalating dosesof the drug until the maximum tolerated dose (MTD) is determined. TheMTD is defined as the dose preceding that at which 2 of 3 or 2 of 6patients experience dose-limiting toxicity.

Test 2:

This is a randomized, multicenter study. The study length is 60 days.Patients are randomized to 1 of 18 treatment groups. For group 1,patients are given salmeterol alone daily at MTD. For Group 2, patientsare given salmeterol on every other day. For Group 3, patients are givensalmeterol once a week. Groups 4-6 have the same dosing regime as 1-3except the dosage is at one-fourth MTD. Groups 7-9 also have the samedosing regime as 1-3 except the dosage is at one-tenth MTD. In additionto the treatment groups, a control group is left untreated.

Endpoint Assessment:

Patients are assessed for reduction of subcutaneous cheek fat ordecrease of buccal fat volume at the conclusion of the study.

Example 6: Effects of a Lipophilic Long-Acting Selective Beta Agonistwhen Injected SC in the Rat Inguinal Fat Pad

The effects of salmeterol xinafoate on the inguinal fat pad weight ofrats were evaluated following their SC injection into the inguinal fatpad. Mature male Sprague-Dawley rats weighing approximately 500 g werelightly anesthetized with 4% isoflurane. Test article injections (drugvehicle, different doses of salmeterol xinafoate) were injected SC intothe right inguinal fat pad in a volume of 0.4 mL. An equal volume ofdrug vehicle was injected into the left inguinal fat pad. Rats receivedan injection comprising doses of 0.1 μs, 1 μg, 10 μg, or 100 μg in eachfat pad on 3 consecutive days. Approximately 24 hours after the lastinjection, rats were anesthetized, sacrificed and the right and left fatpads harvested and weighed. The change in fat pad weight (Right vs.Left) was calculated and expressed as a percent change in inguinal fatpad weight (% change IFP=(Right side−Left side)×100/Left side).

The injection of salmeterol xinafoate into the rat inguinal fat padproduced a dose-related reduction in fat pad weight (FIG. 1). Stainingand histological evaluation of the tissue showed a reduction in fat cellnumber and size (data not shown).

Example 7: Effects of a Lipophilic Long-Acting Selective Beta Agonist onSubcutaneous Fat Thickness in Male and Female Yucatan Minipigs

A study was conducted using 2D-ultrasound to determine the effects ofsubcutaneously administered salmeterol xinafoate (SX) at 0.05, 0.5 or 5μg on subcutaneous fat thickness in male and female Yucatan minipigs.The study was composed of (a control and three treated groups) eachconsisting of 1 minipig/sex for a total of eight minipigs. Each minipighad 4 sites designated for ultrasound measurement; the sites were in a 4cm square orientation along the scapular region. Of the 4 sites, onesite served as an ultrasound control (fat at this site was measuredusing ultrasound, but the site was never injected), while the remaining3 sites were handled as follows: for the control pigs, the 3 remainingsites received an injection of saline during the dosing cycle. In thetreated groups, one site served as a saline control, and 2 sites weredesignated for injection of 0.05, 0.5 or 5 μg of salmeterol xinafoate.Injections were delivered twice per week at a total volume of 1mL/injection site. Each dosing cycle was two weeks in duration. SX wasformulated in a vehicle of 5% PEG400 and 0.25% Tween 80 in water forinjection (WFI).

Ultrasound measurements were made at each treatment site prior to doseadministration on each treatment day, approximately 3-4 days after thelast injection of the dosing cycle, and on the day of necropsy. Bodyweights were collected on the day prior to first dosing and weeklythereafter.

Evaluation of the injection sites did not reveal any adverse reactions.Overall, all animals gained weight over the course of the study (bodyweight increases ranged from 3-7% in males and 6-12% in females). Asseen in FIG. 2, administration of SX produced dose-related changes infat thickness. The lowest doses of SX (0.05 and 0.5 μg) reduced totalfat thickness by approximately 7-9%. The highest dose of SX (5 μg)increased fat thickness slightly (by approximately 3%), possibly due to32-adrenergic receptor tachyphylaxis/desensitization at this dose anddosing schedule.

Example 8: Study of Pharmacokinetics and Bioavailability of LipophilicLong-Acting Selective Beta Agonist in Minipigs

Plasma concentrations of salmeterol were monitored in a male and afemale Gottingen minipig after SC administration of salmeterolxinafoate. SC dose of 4 μg/kg salmeterol xinafoate was administered as asingle bolus dose. Each dose was separated by at least a 3-day washoutperiod. Blood samples (approximately 4 mL) were collected via thebrachiocephalic plexus at pre-dose and at 2, 5, 10, 20, 30, and 45 min,and at 1, 2, 4, 8, and 24 hr post-dose. Blood samples were placed intubes containing K2-EDTA as the anticoagulant. Samples were processed toplasma by centrifugation under refrigeration and stored frozen atapproximately −70° C. (±15° C.) until analysis.

Sample Analysis: Plasma samples were analyzed for salmeterol usingqualified liquid chromatography/mass spectrometry/mass mass spectrometry(LC/MS/MS) methods. The lower limit of quantitation (LLOQ) was 2.50pg/mL for salmeterol.

Data Analysis: Noncompartmental pharmacokinetic parameters werecalculated using WinNonlin 5.2 software, NCA model 202, IV infusioninput for the IV data and NCA model 200, extravascular input for the SCdata (Pharsight Corporation, Mountain View, Calif.). Individual plasmaconcentrations for each animal were used in the calculation ofpharmacokinetic parameters. Nominal collection times and doses were usedin the calculations. The area under the plasma concentration-time curve(AUC) was calculated using linear trapezoidal approximation (linear/loginterpolation). Concentration values below the assay limit ofquantitation were set to zero for calculations. The maximum plasmaconcentration (C_(max)) and the time of its occurrence (T_(max)) wereverified by inspection. The half-life (t_(1/2)) values were calculatedusing the last 3 plasma concentrations with nonzero values, if datapermitted. Other calculations were done using Microsoft Excel® XP. Thebioanalytical and pharmacokinetic data were reported to 3 significantfigures.

Results: The determined pharmacokinetic parameters are shown in Table 4below.

TABLE 4 Pharmacokinetic parameters V_(ss) Dose T_(max) C_(max) AUC_(inf)t_(1/2) t_(last) CL (mL/ Analyte Route (μg/kg) Sex (hr) (pg/mL) (pg ·hr/mL) (hr) (hr) (mL/hr/kg) kg) F Salmeterol SC 4 Fe 0.500 433 1670 8.2324.0 NA NA NA Salmeterol SC 4 M 0.333 564 2260 11.4 24.0 NA NA NAAUC_(inf) = area under the curve at infinite time, CL = plasmaclearance, F = bioavailability, Fe = female, M = male, NA = notapplicable, t_(last) = time of last measurable plasma concentration,V_(ss) = volume of distribution at steady state. SC for fluticasone =single site SC injection, SC for salmeterol xinafoate = single site SCinjection. Male = animal 1-0771, female = animal 2-0623

There were no apparent (consistent) gender differences in the SC PK ofsalmeterol xinafoate. Tmax values ranged from 0.333 to 0.5 hours,indicating that salmeterol xinafoate is rapidly absorbed following SCadministration.

The examples and embodiments described herein are for illustrativepurposes only and various modifications or changes are included withinthe spirit and purview of this application and scope of the appendedclaims. All publications, patents, and patent applications cited hereinare hereby incorporated by reference for all purposes.

What is claimed is:
 1. An injectable monotherapeutic formulation fortreating a body contour defect in a human subject, wherein the singleactive agent is a lipophilic long-acting selective beta-2 adrenergicreceptor agonist, or a salt, solvate, or polymorph thereof that ispresent in an effective amount that is less than about 1 microgram andwherein the formulation is formulated for subcutaneous administrationand further consists of one or more subcutaneously acceptable inactiveingredients.
 2. An injectable monotherapeutic formulation for treating abody contour defect in a human subject, wherein the single active agentis a lipophilic long acting selective beta-2 adrenergic receptoragonist, or a salt, solvate, or polymorph thereof that is present in aneffective amount that is less than about 10 micrograms/milliliter andwherein the formulation is formulated for subcutaneous administrationand further consists of one or more subcutaneously acceptable inactiveingredients.
 3. A method for regional adiposity reduction by amonotherapy treatment regimen, comprising subcutaneously administering amonotherapeutic formulation, wherein each administration during themonotherapy treatment regimen comprises the single active agent selectedfrom a lipophilic long-acting selective beta-2 adrenergic receptoragonist, or a salt, solvate, or polymorph thereof that is present in aneffective amount that is equal to or less than about 20 micrograms, andwherein the formulation further consists of one or more subcutaneouslyacceptable inactive ingredients.
 4. The formulation of claim 1 whereinthe lipophilic long-acting selective beta-2 adrenergic receptor agonistis salmeterol, formoterol, salts, solvents, polymorphs, or a combinationthereof.
 5. The formulation of claim 4 wherein the lipophiliclong-acting selective beta-2 adrenergic receptor agonist is salmeterolxinafoate.
 6. The formulation of claim 5 wherein the formulation isformulated into a weekly dose of salmeterol xinafoate in an amount thatis between about 12 nanograms to about 1 microgram.
 7. The formulationof claim 1 wherein the lipophilic long-acting selective beta-2adrenergic receptor agonist, or a salt, solvate, or polymorph thereofselectively partitions into adipose tissue relative to blood plasma whenadministered to the human subject.
 8. The formulation of claim 1 thatprovides a partition ratio of between 0.01 to about 0.4 when thelipophilic long-acting selective beta-2 adrenergic receptor agonist, ora salt, solvate, or polymorph thereof is administered subcutaneously. 9.The formulation of claim 5 wherein the formulation is formulated to beadministered to the human subject once per week at a single session doseof salmeterol xinafoate that is equal to or less than about 1 microgram.10. The formulation of claim 9 wherein the session dose is divided intoat least two sub-doses of salmeterol xinafoate wherein each sub-dose isin an amount that is between about 12 nanograms to about 1 microgram.11. The formulation of claim 9 wherein the session dose is divided intoat least two sub-doses of salmeterol xinafoate wherein each sub-dose isin an amount that is between about 900 nanograms to about 14 nanograms.12. The formulation of claim 5 wherein the formulation is administeredto the human subject once per week at a session dose of salmeterolxinafoate that is equal to or less than about 650 nanograms.
 13. Theformulation of claim 5 wherein the formulation is administered to thehuman subject in sub-doses of about 5 to about 28 sub-doses.
 14. Theformulation of claim 13 wherein each sub-dose comprises an amount ofsalmeterol xinafoate that is equal to or less than about 950 nanograms.15. The formulation of claim 1 wherein the formulation is formulated tobe administered to a human subject into a submental region, an abdominalregion, a hip region, a thigh region, a buttocks region, a back region,an upper arms region, or a chest region.
 16. The formulation of claim 1wherein the formulation further comprises a subcutaneously acceptableexcipient wherein the excipient is present in an amount of about 0.1milliliter to about 20 milliliters.
 17. The formulation of claim 1wherein the formulation is administered to the human patient in aperiodic dose wherein the periodic dose comprises between 1 and 52 timesper year.
 18. The formulation of claim 1 wherein the formulation is freeof a co-solvent.
 19. The method of claim 3 wherein the method is acosmetic treatment, an aesthetic treatment, or a treatment for thyroideye disease.
 20. The method of claim 3 wherein the lipophiliclong-acting selective beta-2 adrenergic receptor agonist, or a salt,solvate, or polymorph thereof is present in an amount that is betweenabout 980 nanograms and 4 micrograms.